Chicken Meat and Egg Programs - Agrifutures Australia · Sure-feed Pty Ltd PO Box 1208 WAGGA WAGGA...

2000-2001 Research Report

Chicken Meat and Egg Programs

September 2001 RIRDC Publication No 01/073

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© 2001 Rural Industries Research and Development Corporation. All rights reserved.

ISBN 0 642 58295 5 ISSN 1440-6845

“2000-2001 Research Report RIRDC Chicken Meat and Egg Programs” Publication No 01/073 The views expressed and the conclusions reached in this publication are those of the author and not necessarily those of persons consulted. RIRDC shall not be responsible in any way whatsoever to any person who relies in whole or in part on the contents of this report. This publication is copyright. However, RIRDC encourages wide dissemination of its research, providing the Corporation is clearly acknowledged. For any other enquiries concerning reproduction, contact the Communications Manager on phone 02 6272 3186. RIRDC Chicken Meat Program Research Manager RIRDC Egg Program Research Manager Dr Vivien Kite Dr Irene Gorman RIRDC RIRDC PO Box 579 PO Box 569 NORTH SYDNEY NSW 2059 HURSTVILLE NSW 2220 Phone: 02 9929 4077 Phone: 02 9570 9222 Fax: 02 9925 0627 Fax: 02 9570 9763 Email: [email protected] Email: [email protected] RIRDC Publications Manager Rural Industries Research and Development Corporation Level 1, AMA House 42 Macquarie Street BARTON ACT 2600 PO Box 4776 KINGSTON ACT 2604 Phone: 02 6272 3186 Fax: 02 6272 5877 Email: [email protected] Website: http://www.rirdc.gov.au

Published in September 2001 Printed on environmentally friendly paper by Union Offset, Canberra

mailto:[email protected]

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Foreword This year RIRDC has produced Research in Progress, June 2001, which contains short summaries of continuing projects as well as those that were completed during 2000-2001 for all of the Corporation’s 20 program areas. The complete report on all the programs is only available in electronic format on our website at www.rirdc.gov.au. The following report is a hardcopy extract covering Sub-Programs 3.1 and 3.2. It contains all entries from continuing and completed Chicken Meat and Egg research projects funded by RIRDC. Additional information on other activities funded by these programs in 2000-2001 and projects and activities to be funded in 2001-2002 have also been included in this publication. The objective of the Chicken Meat Program is to support increased sustainability and profitability in the chicken meat industry by focussing on research and development on those areas which will enable the industry to become more efficient and globally competitive and which will assist in the development of good industry and product images. The objective of the Egg Program is to support improved efficiency, sustainability, product quality, education and technology transfer in the Australian egg industry. Research reported upon herein was funded from industry revenue which is matched by funds provided by the Federal Government. This report is the newest addition to our extensive catalogue of more than 600 research report, videos and CD-roms of projects supported by RIRDC. Most of our publications are available for viewing, downloading or purchasing online through our website: • downloads at www.rirdc.gov.au/reports/Index.htm • purchases at www.rirdc.gov.au/eshop Peter Core Managing Director Rural Industries Research and Development Corporation

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CHICKEN MEAT PROGRAM ADVISORY COMMITTEE

Chairperson: Mr Barry Shay Executive Director Meat Branch, Safe Food Production NSW PO Box A2613 SYDNEY SOUTH NSW 1235 Ph: (02) 9295 5777 Fax: (02) 9261 2434 Email: [email protected]

Research Manager: Dr Vivien Kite RIRDC Chicken Meat Program PO Box 579 NORTH SYDNEY NSW 2059 Ph: (02) 9929 4077 Fax: (02) 9929 0627 Email: [email protected]

Committee Members: Mr Ian Farran Agribiz Engineering PO Box 279 GEELONG VIC 3220 Ph: (03) 5229 7300 Fax: (03) 5229 7566 Email: [email protected]

Dr Tom Grimes Grimes Consultancy Pty Ltd 4 Henry Street LEWISHAM NSW 2049 Ph: (02) 9569 7436 Fax: (02) 9569 4183 Email: [email protected]

Dr Ron MacAlpine Inghams Enterprises Pty Ltd PO Box 4 LIVERPOOL NSW 2170 Ph: (02) 9606 5666 Fax: (02) 9606 6640 Email: [email protected]

Dr Margaret MacKenzie Inghams Enterprises Pty Ltd PO Box 1100 BROWNS PLAINS QLD 4118 Ph: (07) 3297 0222 Fax: (07) 3297 0578 Email: [email protected]

Dr Harvey Westbury CSIRO Animal Health Private Bag 24 GEELONG VIC 3220 Ph: (03) 5227 5115 Fax: (03) 5227 5555 Email: [email protected]

Dr Jeff Davis RIRDC PO Box 4776 KINGSTON ACT 2604 Ph: (02) 6272 4152 Fax: (02) 6272 5877 Email: [email protected]

http://www.rirdc.gov.au/

http://www.rirdc.gov.au/reports/Index.htm








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EGG PROGRAM ADVISORY COMMITTEE

Chairperson: Dr Andrew Turner Andrew Turner Consulting Pty Ltd Glen Lee 25 Garton Street PRINCES HILL VIC 3054 Ph: (03) 9380 1652 Fax: (03) 9388 8742 Email: [email protected]

Research Manager: Dr Irene Gorman RIRDC Egg Program PO Box 569 HURSTVILLE NSW 2220 Ph: (02) 9570 9222 Fax: (02) 9570 9763 Email: [email protected]

Committee Members: Dr Clive Jackson Biological Technology Transfer Pty Ltd 2 Victory Avenue CAMDEN NSW 2570 Ph: (02) 4655 4007 Fax: (02) 4655 4008 Email: [email protected]

Mr Noel Kratzmann DA Hall & Co PO Box 49 MILLMERRAN QLD 4357 Ph: (07) 4695 1717 Fax: (07) 4695 1717 Email [email protected]

Mr Geoff Munzberg Munzberg & Co. Pty Ltd PO Box 166 TANUNDA SA 5352 Ph: (08) 8563 2625 Fax: (08) 8563 2027 Email: [email protected]

Ms Judith O’Keeffe Sure-feed Pty Ltd PO Box 1208 WAGGA WAGGA NSW 2650 Ph: (02) 6925 6066 Fax: (02) 6926 5960 Email: [email protected]

Dr Peter Scott Scolexia Pty Ltd 16 Learmonth Street MOONEE PONDS VIC 3039 Ph: (03) 9326 0106 Fax: (03) 9372 7576 Email: [email protected]

Mr Philip Szepe Kinross Farm Pty Ltd 2110 Yea Road KINGLAKE WEST VIC 3757 Ph: (03) 5780 1242 Fax: (03) 5780 1238 Email: [email protected]

Dr Jeff Davis RIRDC, PO Box 4776 KINGSTON ACT 2604 Ph: (02) 6272 4152 Fax: (02) 6272 5877 Email: [email protected]

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INDEX TO PROJECT SUMMARIES

CHICKEN MEAT PROGRAM - COMPLETED PROJECTS

Flock Health BTT-2A* Revision of the AUSVETPLAN Disease Strategy document on very virulent Infectious

Bursal Disease ....................................................................................................................... 1 CSA-4J* Postgraduate Scholarship – Matthew Rudd: Identification of virulence determinants of

infectious bursal disease virus (IBDV).................................................................................... 3 CSA-7J* Therapeutic applications of chicken interferon gamma in poultry .......................................... 5 DAN-159J* NDV vaccination strategies aiming to induce high HI titres in elite breeding and layer

flocks ...................................................................................................................................... 7 DAQ-244J* Investigations into the management of the darkling beetle, Alphitobius diaperinus

(Panzer).................................................................................................................................. 8 DAW-98A The efficient production of big liver and spleen vaccine antigen.......................................... 10 RMI-6J* The development of effective immunisation strategies against Marek’s disease ................ 11 RMI-8J* Characterisation of very virulent Australian isolates of Marek’s disease virus .................... 13 UMO-21A Candidate vaccine antigens against Pasteurella multocida ................................................. 15 UNE-70A Effects of enzymes on gut microbial status in broiler chickens............................................ 17 UTS-3J* Postgraduate scholarship – David Witcombe: Production and characterisation of

recombinant antigens of Eimeria.......................................................................................... 18

Bird Nutrition and Feed Supply GRD-1J* Premium Grains for Livestock Program ............................................................................... 20 UNE-64A Postgraduate scholarship- Andreas Kocher: Increasing the nutritive value of grain

legumes for poultry by use of more efficacious enzyme systems........................................ 24 US-68A Postgraduate scholarship – Ron Newman: Manipulation of lean tissue deposition in

broiler chickens by altering the sensitivity of tissues to insulin ............................................ 27

Food Safety DAQ-245A Risk factors for Campylobacter spp. in Australian broilers .................................................. 29 IMVS-1A Molecular basis of benign colonisation of Salmonella Sofia in chickens ............................. 31 RMI-7A Use of avirulent Campylobacter jejuni strains to control poultry-derived campylobacter

food poisoning ...................................................................................................................... 33 USA-9A Antibiotic resistance in bacteria isolated from poultry .......................................................... 36

Animal Welfare DAV-162A A welfare audit for the transport and processor sectors of the broiler industry.................... 38

Environmental Management DAW-94A Conditioning and Analysis of Broiler litter to prevent fly breeding........................................ 41

Other DAN-142A Production of Trial Distance Learning Materials for the Poultry Industry - Stage III ............ 43

* Indicates a joint Chicken Meat and Egg Program project. These projects will appear in both sections of this report.







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CHICKEN MEAT PROGRAM - RESEARCH IN PROGRESS

Flock Health CSA-1J* Detection of virulent strains of Newcastle disease virus in chickens previously infected

with Australian strains of the virus........................................................................................ 45 CSA-10J* Postgraduate scholarship – Ms Louise Hilton: Therapeutic applications of cytokines in

poultry................................................................................................................................... 46 CSA-11J* Molecular epidemiology of Newcastle disease virus in Australia......................................... 47 CSA-12A Biological control of necrotic enteritis in meat chickens....................................................... 48 CSA-13J* Postgraduate Scholarship - Jacqueline Kattenbelt: Mapping of structure-function

relationships of Newcastle Disease (ND) using reverse genetics........................................ 49 CSA-15J* Diagnostic tools for differentiation of vvIBDV and characterisation of Australian strains..... 50 CSA-16A Evaluation of fowlpox (FPV) strains free of reticuloendotheliosis virus (REV) as vaccines

for use in Australian poultry flocks........................................................................................ 51 DAN-171A Infectious proventriculitis and stunting syndrome of broiler chickens .................................. 52 DAQ-259J* Attenuation and characterisation of chicken Eimeria for live vaccines ................................ 53 DAQ-273A Investigations into the development of a sustainable management strategy for the

darkling beetle, Alphitobius diaperinus (Panzer) in broilers ................................................. 54 MS990-40* National NDV Survey ........................................................................................................... 55 RMI-11A The development of vaccination strategies to control necrotic enteritis in poultry............... 57 UM-45J* Determination of the genomic sequence of Mycoplasma gallisepticum .............................. 58 UM-49A Avian Leukosis-J (ALV-J) in Australia: laboratory technologies and research needs.......... 59 UMU-23J* Control of intestinal spirochaete infections in chickens........................................................ 60 UNE-75A Effects of organic acids, prebiotics, and enzymes on control of necrotic enteritis and

performance of broiler chickens ........................................................................................... 61 US-72J* Enhancing mucosal immunity in chickens by novel in-ovo and postnatal vaccination

techniques ............................................................................................................................ 62

Bird Nutrition and Feed Supply DAQ-264J* Characterisation of canola meal and cottonseed meal at practical inclusion levels for use

in broiler and layer diets ....................................................................................................... 63 DAQ-277A Estimating lysine availability by slope-ratio chick assay ...................................................... 65 GRD-2J* Inclusion of data for additional livestock species in the Australasian Livestock Feed

Ingredient (ALFI) database................................................................................................... 66 GRD-3J* Premium Grains for Livestock Program (stage 2) ................................................................ 67 SAR-13A Physiological limitations in energy metabolism reduce production efficiency of broilers ..... 68 US-80A Improving the utilisation of dietary amino acids in meat chickens ....................................... 69 US-104A Use of dietary fatty acids to increase protein accretion in broilers....................................... 70

Food Safety IMV-3A Salmonella typing and colonisation of chickens by characterised S. Sofia.......................... 71 UG-3A Development of campylobacter bio-replacement program and establishment of

campylobacter reference centre........................................................................................... 73

Animal Welfare DAV-185A Implementation of the RIRDC broiler welfare audit to industry ............................................ 75

Environmental Management JSC-1A Sustainability improvements in the Victorian chicken meat industry (phase 1) ................... 76 SAR-33J* Reduction of dust emissions from broiler and caged layer sheds........................................ 78


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EGG PROGRAM - COMPLETED PROJECTS

Implications of the Changing Economic Environment for the Australian Egg Industry AEI-8A/ National industry databases ................................................................................................. 112H79 AEI-9A IMS-3A Review of information sources used by Australian rural industries and egg industries

overseas ............................................................................................................................... 113H81

Flock Health and Disease Management BIR-1A Layer industry disease and management survey................................................................. 114H83 BTT-2A* Revision of the AUSVETPLAN Disease Strategy document on very virulent Infectious

Bursal Disease ..................................................................................................................... 115H86 CSA-4J* Postgraduate Scholarship – Matthew Rudd: Identification of virulence determinants of

infectious bursal disease virus (IBDV).................................................................................. 116H88 CSA-7J* Therapeutic applications of chicken interferon gamma in poultry ........................................ 117H90 DAN-159J* NDV vaccination strategies aiming to induce high HI titres in elite breeding and layer

flocks .................................................................................................................................... 118H92 DAQ-244J* Investigations into the management of the darkling beetle, Alphitobius diaperinus

(Panzer)................................................................................................................................ 119H93 RMI-6J* The development of effective immunisation strategies against Marek’s disease ................ 120H95 RMI-8J* Characterisation of very virulent Australian isolates of Marek’s disease virus .................... 121H97 UM-37A Development of a live attenuated vaccine for chicken anaemia virus ................................. 122H99 UTS-3J* Postgraduate scholarship – David Witcombe: Production and characterisation of

recombinant antigens of Eimeria........................................................................................ 123H101

Feed Availability and Nutrition DAQ-241A Alternative protein sources for laying hens ........................................................................ 124H103 GRD-1J* Premium Grains for Livestock Program ............................................................................. 125H106 US-54A Amino acid and energy requirements of imported IsaBrown laying hens.......................... 126H110

Husbandry and Welfare US-71A Development of a non-invasive test of stress in laying hens ............................................. 127H112

Environmentally Sustainable Management UNE-59A Environmentally acceptable land use for chicken meat and egg production ..................... 128H113

EGG PROGRAM - RESEARCH IN PROGRESS

Implications of the Changing Economic Environment for the Australian Egg Industry ANU-41A The economic impact of changing Australian egg production systems ............................. 129H115 AEI-11A Options for enhancing industry competitiveness and R&D and marketing efficiency

(EIDF) ................................................................................................................................. 130H117

New and Existing Markets DAQ-275A An evaluation of the higher value-added opportunities from the chicken egg ................... 131H118


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Public Health CIF-1A Rapid detection of virulent Salmonella in egg and poultry products .................................. 119 UNE-71A Egg and egg shell quality control in the Australian egg industry........................................ 133H120

Flock Health and Disease Management AEI-10A Trialing emergency animal disease arrangements in the Australian egg industry (EIDF) . 134H121 CSA-1J* Detection of virulent strains of Newcastle disease virus in chickens previously infected

with Australian strains of the virus...................................................................................... 135H123 CSA-10J* Postgraduate scholarship – Ms Louise Hilton: Therapeutic applications of cytokines in

poultry................................................................................................................................. 136H124 CSA-11J* Molecular epidemiology of Newcastle disease virus in Australia....................................... 137H125 CSA-13J* Postgraduate Scholarship - Jacqueline Kattenbelt: Mapping of structure-function

relationships of Newcastle Disease (ND) using reverse genetics...................................... 138H126 CSA-15J* Diagnostic tools for differentiation of vvIBDV and characterisation of Australian strains... 139H127 DAQ-259J* Attenuation and characterisation of chicken Eimeria for live vaccines .............................. 140H128 DAV-170A Studies of cloacal haemorrhage, egg peritonitis, vent trauma and beak trimming in the

laying hen ........................................................................................................................... 141H129 MS990-40* National NDV Survey ......................................................................................................... 142H130 UJC-7A Molecular diagnostic tools for wild type and vaccine strains of Marek's disease virus...... 143H132 UM-45J* Determination of the genomic sequence of Mycoplasma gallisepticum ............................ 144H133 UM-51A Investigating sanitation of surface water for poultry using chlorine - IBDV models ........... 145H134 UMU-23J* Control of intestinal spirochaete infections in chickens...................................................... 146H136 UNE-72A Effects of diet composition, gut microbial status and feed forms on cannibalism in

layers .................................................................................................................................. 147H137 UNE-76A Optimising infectious bronchitis vaccination of laying hens for maximum egg shell

quality ................................................................................................................................. 148H138 US-72J* Enhancing mucosal immunity in chickens by novel in-ovo and postnatal vaccination

techniques .......................................................................................................................... 149H139

Feed Availability and Nutrition DAQ-264J* Characterisation of canola meal and cottonseed meal at practical inclusion levels for use in

broiler and layer diets ......................................................................................................... 150H140 GRD-2J* Inclusion of data for additional livestock species in the Australasian Livestock Feed

Ingredient (ALFI) database................................................................................................. 151H142 GRD-3J* Premium Grains for Livestock Program (stage 2) .............................................................. 152H143 UNC-12A Hind gut function in laying hens ......................................................................................... 153H144 UNE-77A Effects of commercial feed enzymes in wheat-based diets on egg and egg shell quality

in imported strains of laying hen......................................................................................... 154H145 UWA-61A Evaluation of Lathyrus cicera as a feed ingredient for layers ............................................ 155H146

Husbandry and Welfare SAR-34A Should claw abrasives be used in cages in Australia? ...................................................... 156H147 SAR-35A Beak trimming accreditation ............................................................................................... 157H148

Environmentally Sustainable Management SAR-33J* Reduction of dust emissions from broiler and caged layer sheds...................................... 158H149 UQ-93A The assessment and development of best management practice techniques for

Australian laying hens housed in conventional and alternative laying systems................. 159H150 UQ-97A Pilot study on the use of time lapse video to study the behaviour of laying hens in

conventional and modified cages....................................................................................... 150 * Indicates a joint Chicken Meat and Egg Program project. These projects will appear in both sections of this report.

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Training, Information and Technology Transfer DAN-138A National egg industry newsletter ........................................................................................ 160H153 DAN-189A Video series - Workplace training for layer farm staff ........................................................ 161H154 SAR-36A Vaccination training manual ............................................................................................... 162H155

OTHER SUPPORTED ACTIVITIES ............................................................................ 163H156

SCHOLARSHIPS...................................................................................................................... 164H156

RESOURCE DEVELOPMENT.................................................................................................. 165H156

TRAVEL/CONFERENCE/WORKSHOPS................................................................................. 166H157

RESEARCH TO BE SUPPORTED IN 2001/2002 CHICKEN MEAT PROGRAM....... 167H158

RESEARCH TO BE SUPPORTED IN 2001/2002 EGG PROGRAM .......................... 168H160 * Indicates a joint Chicken Meat and Egg Program project. These projects will appear in both sections of this report.

CHICKEN MEAT PROGRAM – COMPLETED PROJECTS 1

CHICKEN MEAT PROGRAM

COMPLETED PROJECTS

Flock Health

Project Title

Revision of the AUSVETPLAN Disease Strategy document on very virulent Infectious Bursal Disease

RIRDC Project No:

BTT-2A

Researcher: Dr Clive Jackson Organisation: Biological Technology Transfer

2 Victory Avenue CAMDEN NSW 2570

Phone: (02) 4655 4007 Fax: (02) 4655 4008 Email: [email protected]

Objective

• To revise the AUSVETPLAN Disease Strategy document on very virulent

infectious bursal disease.

Background

In 1998, Dr Jackson developed an AUSVETPLAN Disease Strategy Document as part of a consultancy to the DPIE. That Strategy Document was developed because of the serious economic impact that very virulent Infectious Bursal Disease (vvIBD) would have on the poultry industry should it gain entry into Australia. The potential annual loss was estimated to be in the order of $50 million. A revised Strategy Document was developed using the Newcastle Disease (ND) Strategy Document as a model. However, experiences gained through the implementation of that document in the course of the 1998-2000 outbreak of virulent ND in NSW resulted in the need for further revision of the Disease Strategy document for vvIBD.

Research The revision was undertaken with the assistance of a corresponding committee of Australian scientific experts on IBD. The Strategy Document was rewritten to incorporate decisions from a government/industry meeting held on 19 August 1999. The revised Strategy Document also considered the role played by government and industry during the 1998-2000 eradication of virulent Newcastle disease in NSW. It also considered the new cost-sharing agreement for exotic disease control and eradication.

Outcomes A revised Disease Strategy document has been developed for review by RIRDC Egg and Chicken Meat Programs and industry. The revised Strategy Document should provide the industry and government with a contemporary document on which to base an eradication program. The Strategy Document takes into account the current


funding arrangements for exotic disease control in Australia. It attempts to define the resources required by industry and government. It also provides industry with guidance in undertaking a risk analysis of the benefits to be derived from pursuing eradication with limited resources and funding.

Implications The existence of a Disease Strategy document focuses the attention of the industry on the need to remain vigilant against the entry of vvIBDV. It emphasises the need to continue to develop rapid diagnostic tests to detect the presence of any very virulent viruses as early as possible. It also demands the availability of funds and resources to undertake any eradication program.

Publications Jackson, C.A.W. (2001) AUSVETPLAN Disease Strategy for very virulent infectious bursal disease virus (vvIBDV) eradication. Proc. AVPA Scientific Meeting 6-7.2.01 University of Sydney, p15.

Jackson, C.A.W. (2001) A Disease Strategy for the prevention and eradication of very virulent infectious bursal disease virus (vvIBDV). Proc. XIIth International Congress of the WVPA, Cairo, 17-21.9.01 (Abstract) (Submitted for publication).



Project Title

Postgraduate Scholarship – Matthew Rudd: Identification of virulence determinants of infectious bursal disease virus (IBDV)

RIRDC Project No:

CSA-4J

Researcher: Mr Matthew Rudd and Dr Jagoda Ignjatovic Organisation: CSIRO Livestock Industries

Australian Animal Health Laboratory (AAHL) Private Bag 24 GEELONG VIC 3213


Objectives

• To characterise and sequence a very virulent (vv) exotic strain of infectious

bursal disease virus (IBDV) and identify genomic region(s) which may influence pathogenesis.

• To compare and contrast the pathogenicity of attenuated and very virulent IBDV (vvIBDV) strains in vivo, and identify criteria which might differentiate between the two strains.

• To establish a reverse genetics system for the recovery of infectious IBDV from cell culture, embryonating eggs and/or directly from SPF chickens.

• To swap genomic material between attenuated and vvIBDV strains and assess the impact of such manipulations on pathogenicity in vivo.

Background

Since first reported in 1989, vvIBDV has spread rapidly throughout Europe, Asia, and many other countries. Australia is currently free of such strains. This project seeks to identify potential virulence markers of IBDV for the development of rapid and highly accurate diagnostic tests which could be used to detect any incursion of exotic vvIBDV strains, should this ever occur.

Research An Indonesian vvIBDV strain was completely sequenced and the deduced amino acid sequences were aligned with the corresponding sequences of published IBDV strains to identify amino acids which are conserved solely in very virulent strains. An endemic attenuated IBDV strain (002-73) and an Indonesian vvIBDV strain (Tasik94) were both extensively characterised in vivo, and the ability of several biological assays to differentiate between the two strains were assessed. Genomic material, corresponding to a portion of the VP2 protein, was swapped between the attenuated 002-73 and very virulent Indonesian Tasik94 strains to generate recombinant IBDV. A reverse genetics system will be used to recover infectious recombinant virus. Pathogenicity testing of the recombinant virus will be used to assess the significance of putative virulence determinants identified and to provide valuable information which will assist in the development of a diagnostic assay.

Implications Further work needs to be conducted to recover and assess recombinant virus(es). The successful identification of virulence determinants is a prerequisite for


developing diagnostic tests needed to monitor the field situation of IBDV in Australia.

Publications Rudd, M.F., Heine, H.G., Parede, L., Sapats, S.I., Ignjatovic, J. (2001) Characterisation of an Indonesian strain of very virulent infectious bursal disease virus (vvIBDV). Proc. Int. Symp. Infectious Bursal Disease Virus and Chicken Anemia. In press.

Currently in preparation: Rudd, M.F., Heine, H.G., Middleton, D., Lowther, S., Ignjatovic, J. Differentiation

between endemic and exotic strains of infectious bursal disease virus (IBDV). Poster in preparation for presentation at the 3rd Veterinary Virology Conference, 26-28th September 2001.

Rudd, M.F., Heine, H.G., Sapats, S.I., Ignjatovic, J. Identification of putative virulence determinants of infectious bursal disease virus (IBDV). Manuscript in preparation for submission to Virus Research.


Project Title

Therapeutic applications of chicken interferon gamma in poultry

RIRDC Project No:

CSA-7J

Researcher: Dr John Lowenthal Organisation: CSIRO Livestock Industries



Objective

• To enhance disease resistance and vaccine efficacy in poultry by administering

therapeutic doses of chicken interferon-gamma to commercial broilers and layers.

Background

Safe, natural alternatives to antibiotics for the maintenance of optimal growth rates and flock health in chickens are being sought by the poultry industry worldwide. Cytokines are natural proteins that are produced by the body’s immune system during infection. Their ability to protect against disease make them excellent candidates as naturally occurring therapeutics.

Research Previous studies on chicken interferon gamma (ChIFN-γ) identified it as having therapeutic potential. In this project, the ability of ChIFN-γ to act as a growth promoter and vaccine adjuvant was assessed in trials using broiler chickens under commercial conditions.

Outcomes An Escherichia coli expression system was developed for the large scale production of recombinant ChIFN-γ protein. The recombinant ChIFN-γ was found to be biologically active. Monoclonal antibodies were produced and an ELISA was developed for the detection of ChIFN-γ. Treatment of broilers with ChIFN-γ resulted in enhanced weight gain over a period of up to eight weeks compared to control birds. ChIFN-γ treatment also reduced weight loss suffered by birds following infection with Eimeria acervulina.

Implications Rapid transfer of this technology to the Australian poultry industry is anticipated. These results show the feasibility of using cytokines as natural therapeutics and adjuvants. Additional cytokines have recently been identified, including interleukins -2, -6, -15 and -18. These are now available for a similar type of assessment. The particular activities of these new cytokines make them attractive new treatments for diseases such as coccidiosis, Marek’s disease and infectious bronchitis.

Publications Lowenthal, J.W., O'Neil, T.E., Strom, A.D.G., and Andrew, M.E. (1999) Cytokine therapy: a natural alternative for disease control. Vet. Immunol. Immunopathol. 72, 183-188.


Lowenthal, J.W., Lambrecht, B, van den Berg, T.P., Andrew, M.E., Strom, A.D.G., and Bean, A.G.D. (2000) Avian cytokines – the natural approach to therapeutics. Dev. Comp. Immunol. 24, 355-365.

Lowenthal, J.W. (2001) Therapeutic applications of cytokines - what can the chicken teach us? Avian Dis. (in press).

Lowenthal, J.W., Richards, G.G., Bean A.D.G., O’Neil T.E., Hilton L.S., Tyac S., Pooley, C., and Johnson M.A. (2001) New vaccination strategies for control of coccidiosis. Avian Dis. (in press).

Hilton, L.S., Bean, A,G.D., and Lowenthal, J.W. (2001) Recent advances in avian cytokines. Vet. Immunol. Immunopatol. (Invited Review – in press).

Lowenthal, J.W. et al. Chicken interferons: natural therapeutics in poultry. Avian Immunology Research Group Meeting, Turku, Finland 1998.

Lowenthal, J.W. Cytokine therapy – a natural alternative for disease control. 4th Asia Pacific Health Conference, Melbourne, 1998.

Lowenthal, J.W. Cytokine therapy – a natural alternative for disease control. 5th International Veterinary Immunology Symposium, India, 1998.

Lowenthal, J.W. Practical applications of chicken cytokines. 11th Australian Poultry and Feed Convention, Gold Coast. 1999.

Lowenthal, J.W. et al. New strategies for enhancing immunity to Eimeria: interferon gamma is the natural approach to disease control. Vaccines against Coccidioses conference, Dublin, 2000.

Lowenthal, J.W. Therapeutic applications of cytokines - what can the chicken teach us? Avian Immunology Research Meeting, Cornell University, USA, 2000.

Lowenthal, J.W. et al. New vaccination strategies for control of coccidiosis. Avian Immunology Research Meeting, Cornell University, USA, 2000.

Lowenthal, J.W. et al., Cytokines are the next generation of therapeutics. AVAP Meeting, Attwood, Vic. Oct 2000.


Project Title

NDV vaccination strategies aiming to induce high HI titres in elite breeding and layer flocks

RIRDC Project No:

DAN-159J

Researcher: Dr George Arzey Organisation: NSW Department of Agriculture

Elizabeth Macarthur Agricultural Institute (EMAI) PMB 8 CAMDEN NSW 2570


Objective

• To identify strategies capable of producing the highest and most persistent mean

HI titres in vaccinated flocks.

Background

The recent availability of inactivated ND vaccines in Australia has broadened the scope for effective long-term protection of birds against Newcastle disease. However, no sound data was previously available on the HI titres elicited when V4 was used as the priming vaccine. The present study was undertaken in order to assess a range of vaccination strategies in caged layers with the use of V4 as the priming agent

Research Ten different vaccination strategies were evaluated by monitoring the NDV HI response over a period of 28 weeks in serologically negative 18 weeks old pullets housed in commercial cages.

Outcomes Any of the combinations of live V4 plus inactivated vaccine trialled should protect a flock against clinical Newcastle disease. For protection against a drop in egg production, the strategies in which V4 was followed by inactivated vaccine four weeks later can be considered protective up to 24 weeks post initial vaccination. Protection against infection for up to three months (as determined by reduced shedding of a virulent virus) can be expected with V4 delivered orally followed with inactivated vaccine four weeks later. The studies also confirmed the poor ability of V4 to spread in flocks.

Implications The study demonstrated that V4 used as a priming vaccine with a selective combination of an inactivated vaccine can produce high and persistent NDV HI titres that are comparable to those elicited by other live overseas vaccines in combination with inactivated vaccines.


Project Title

Investigations into the management of the darkling beetle, Alphitobius diaperinus (Panzer)

RIRDC Project No:

DAQ-244J

Researchers: Mr Trevor Lambkin and Mr MC Cameron Organisation: Department of Primary Industries (Qld)

Farming Systems Institute 80 Meiers Rd INDOOROOPILLY QLD 4068

Phone: (07) 3896 9434 Fax: (07) 3896 9446 Email: [email protected], [email protected]

Objectives

• To review the relevant literature pertaining to A. diaperinus research and

thereby develop an improved understanding of the ecology of the pest. • To develop an insecticide resistance testing method for A. diaperinus and

subsequently survey and test broiler shed and egg barn pest populations in south east Queensland for insecticide resistance.

Background

The darkling beetle, Alphitobius diaperinus (Panzer) is a common cosmopolitan insect pest of poultry houses, in particular broiler sheds and egg barns, and is capable of transmitting a large number of poultry diseases and parasites. In recent concern has been expressed about increasing beetle numbers in broiler sheds and the pest’s potential to breach farm bio-security. In spite of the occurrence of the pest on almost every Australian poultry farm, no previous Australian research has been undertaken to determine the pest’s behaviour and its insecticide resistance status. Research commenced in 1998 to address these gaps in our understanding of the pest; in particular, to develop a better understanding of the ecology of the pest and to determine if resistance to fenitrothion (Folithion®) and cyfluthrin (Tugon®) has occurred in pest populations.

Research A review of scientific literature on darkling beetle research was undertaken to provide improved knowledge of the pest’s ecology, published research results and possible control strategies. Work was undertaken towards the development of an effective and efficient laboratory culture method for A. diaperinus, in order to provide a constant and adequate supply of beetles for laboratory research and testing with insecticides. A laboratory insecticide-resistance testing method was developed to provide the tools necessary to identify and characterise any insecticide resistance in A. diaperinus populations. A survey was undertaken of local broiler shed and egg barn beetle populations and these populations were subsequently tested with fenitrothion (and some with cyfluthrin). Each population tested was compared to a susceptible reference population and from this comparison a level of resistance was determined.

Outcomes The scientific literature review and project field studies have indicated that the pest’s ability to avoid contact with insecticides contributes to A. diaperinus control failures. This behaviour, together with predominantly clay-floored sheds in most broiler sheds contributes to problems with control after clean-outs, as many individuals stay concealed in the floor and do not receive a lethal dose of insecticide. The



development of a laboratory culture method has provided adequate numbers of some test insects. Problems with the culture method arose during the latter part of the project due to mite infestations, and these have hindered the availability of insects for testing. The development of fenitrothion and cyfluthrin resistance testing methods has been successful. Test results have shown that insects from south east Queensland broiler systems have strong fenitrothion resistance and some cyfluthrin resistance, and preliminary results indicate that populations of A. diaperinus from some production areas, for example Armidale and the Atherton Tablelands, have quite weak fenitrothion resistance. Insecticide resistance levels in insects from other intensive livestock systems, including egg barns are generally weaker, and all levels of resistance are directly related to duration and frequency of insecticide use.

Implications Insecticide resistance is not the major factor that determines beetle population sizes in broiler sheds. There is no relationship between anecdotal estimates of broiler beetle numbers and fenitrothion and cyfluthrin resistance levels, ie- population sizes of insects in different broiler sheds, with similar levels of insecticide resistance, can be very different. Results of testing A. diaperinus from the only south east Queensland egg barn studied show that the insects are still susceptible to fenitrothion and a rotation of fenitrothion and cyfluthrin may be done on alternate clean outs. Whether these egg barn results are typical for all is not known. For broiler sheds in general it is suggested that the application of cyfluthrin may be reduced to every second clean out (part or full) or just used over the summer period. This may delay the development of stronger resistance. Preliminary results for the broiler production areas of Armidale and the Atherton Tablelands indicate that fenitrothion may be included with cyfluthrin in an insecticide rotation. In summary, as development of strong insecticide resistance in all areas is inevitable given time, a closer examination is needed of the other major factors that control population sizes in broiler sheds. When this is known, studies of currently registered insecticides, alternative control strategies and the interaction of both can be properly evaluated.

Publications Lambkin, T.A. (1998) Controlling black beetles (Alphitobius diaperinus) in chicken

sheds. Proceedings 1998 Poultry Exchange Surfers Paradise 19-21 April 1998, 33-37.

Lambkin, T.A. and Cameron, M.C. (1999) Darkling beetle control-current difficulties and future prospects. Proceedings 11th Australian Poultry & Feed Convention 10-13 October 2000, 184-192.

Lambkin, T.A. & Cameron, M.C. (2000) Darkling beetle control in Australian broilers - a new direction. Proceedings 2000 Poultry Exchange Surfers Paradise 9-11 April 2000, 97-102.


Project Title

The efficient production of big liver and spleen vaccine antigen

RIRDC Project No:

DAW-98A

Researcher: Dr Sarah Plant Organisation: Agriculture Western Australia

3 Baron Hay Court SOUTH PERTH WA 6151


Objectives

• To assess alternative methods of BLS antigen production. • To reduce the financial losses associated with BLS through the development and

assessment of efficient techniques for the production of BLS vaccine antigen.

Background

Earlier RIRDC-funded projects developed highly specific and sensitive diagnostic tests for big liver and spleen (BLS) infection and identified a candidate vaccine for BLS disease of broiler breeder hens. The vaccine was shown to generate a satisfactory response to BLS and vaccinated birds demonstrated the ability to clear a BLS infection faster than unvaccinated birds. The production of this vaccine is cumbersome and not economically suitable for large scale commercialisation.

Research Two methods of producing a less expensive BLS vaccine were examined. One method used a small protein (peptide) selected for its antigenic similarity to the BLS protein (phage library). This peptide was expressed on a bacteriophage (bacterial virus) which was simple and inexpensive to produce in large quantities. The second method attempted to express the BLS protein in an insect cell culture system. The bacteriophage-expressed protein was inoculated into hens as a synthetic peptide and as expressed on the bacteriophage in a small-scale vaccine trial, where antibody responses were monitored.

Outcomes A peptide was identified which had antigenic similarity with the BLS protein. Hens inoculated with a synthetic preparation of this peptide demonstrated increased antibody titres to the peptide following inoculation. This increase in antibody titre was not observed in hens inoculated with the peptide expressed on the bacteriophage. No BLS protein was produced in the insect cell culture system.

Implications This project showed that the bacteriophage system selected peptides with antigenic similarity to the BLS protein. The vaccine study also showed encouraging antibody responses in the synthetic peptide inoculated birds. Future work should determine if the vaccinated birds demonstrate an improved ability to clear BLS viral infection. The peptide vaccine may represent a more cost-effective means of BLS vaccine production.


Project Title

The development of effective immunisation strategies against Marek’s disease

RIRDC Project No:

RMI-6J

Researcher: Prof Greg Tannock Organisation: RMIT University

Virology Laboratory PO Box 71 BUNDOORA VIC 3083


Objectives

• To improve the performance of existing Marek’s disease vaccines using suitable

adjuvants. • To characterise MDV strains after adaptation to continuous cell lines. • To develop a MDV serotype 1 specific probe to use in a dot-blot hybridisation

technique.

Background

For almost 30 years, Marek’s disease (MD) has been largely controlled by the use of intensive vaccination with herpesvirus of turkeys (HVT), either alone or in combination. However, in recent years HVT vaccines have shown to be much less effective against emerging field strains of MDV of increasing virulence. Adjuvants are used to improve the immunogenicity of a vaccine without increasing the amount of infectious virus in the vaccine. An adjuvant enhancing the performance of the HVT vaccine would be a value-added benefit to a cheap and readily available existing vaccine. The adaptation of MDVs to growth in a continuous cell line could be useful for vaccine production, compared with the labour and reagent intensive CEF cultures that are currently used. Because of limitations associated with current detection techniques for MDV serotype 1 virus, a MDV1 specific probe used in a rapid identification assay which is less expensive and more specific than those currently available, would be very useful for field diagnosis and important in vaccine evaluation.

Research Commercial broiler chickens were used to assess the possible role of γ-inulin as an adjuvant to improve the efficiency of HVT vaccination against MD. Chickens were administered vaccine with or without γ-inulin using three vaccination procedures: (i) in-ovo, by Inovoject®, (ii) in ovo by hand, or (iii) subcutaneously (sc) at day old. All birds were challenged with a virulent MDV 1 challenge virus at three days of age. Effective vaccination by HVT was assessed by the development of viraemia. HVT and MDV1 were adapted to the Vero continuous cell line (Jaikumar, 2001) and were characterised by immunological and molecular techniques. A probe labelled with digoxigenin (DIG) was developed for the detection of MDV 1


by dot-blot hybridisation. The probe was labelled by the incorporation of DIG in a PCR reaction product that consisted of the amplified 132 bp repeat located within the inverted repeat long region of the MDV 1 genome. The sensitivity and specificity of virus isolation and dot-blot hybridisation were compared with PCR, which was used as the reference procedure.

Outcomes In the vaccination trial, MD was present in all treatment groups but its incidence in groups treated with γ-inulin was not significantly different from non-treated groups. Differences in the percentages of MD in groups administered γ-inulin using the Inovoject® method or sc at day 1 were also non-significant. No adverse effects due to γ-inulin were noted in any group. HVT grew more rapidly and produced more extensive CPE and higher virus yields in Vero cell lines than MDV1. When the genome of adapted serotype 1 viruses was examined, an expansion of the 132 bp DR sequence indicated that the infected cell line contained serotype 1 MDV DNA. The presence of intact DNA with a size of approximately 180 kb in both serotype 1- and HVT-infected Vero cells after isolation and characterisation indicated that whole copies of both types of DNA were present and provided further evidence for adaptation to growth of the serotype 1 virus. This is the first report of the growth of either virus in a continuous line. Highest sensitivity rates were achieved by dot-blot hybridisation using the 132 bp PCR probe, compared with virus isolation and identification by immunoperoxidase or immunofluorescence. Despite their much lower sensitivity, higher specificity was obtained by both culture detection methods than for the dot-blot hybridization

Implications This project has shown that γ-inulin did not appear to function as an adjuvant when administered with HVT vaccine. The presence of intact DNA with a size of approximately 180 kb in both serotype 1- and HVT-infected Vero cells after isolation and subsequent analysis in a pulsed-field gel indicates that whole copies of both types of DNA were present and provides further support for adaptation to growth of the serotype 1 virus, thus would be useful for new and improved vaccine production procedures. The dot-blot hybridisation technique using the DIG-labelled probe specific for MDV 1 was shown to be potentially very useful as a rapid and economical test to detect MDV.

Publications Cipriani, T. L. (2000) The development of two digoxigenin-labelled probes for the detection of Marek’s disease virus by dot-blot hybridisation. B. App.Sc.Honours Thesis.

Jaikumar, D. (1998) Propagation and molecular characterisation of Marek’s disease virus. Master of App.Sc. Thesis.

Jaikumar, D. (2001) Adaptation of Marek’s disease virus to the Vero continuous cell line. Veterinary Microbiology 70, 75-82.


Project Title

Characterisation of very virulent Australian isolates of Marek’s disease virus

RIRDC Project No:

RMI-8J




Objectives

• To collect and maintain a repository of serotype 1 strains of Marek’s disease

virus (MDV) representative of Australia and to characterise them for future reference.

• To determine optimum methods for standardising Marek’s disease (MD) vaccines.

• To prepare and maintain reference preparations of NDV for use in vaccine assays.

• To provide an independent, reliable vaccine assay facility for use by Australian vaccine manufacturers in harmony with requirements set by NATA.

• To continually maintain and supply MDV challenge preparations for use by the industry.

Background

MD is currently a significant economic problem for the Australian chicken industry. Current, locally developed vaccines appear to provide very little protection against recently isolated very virulent strains of MDV. The availability of new vaccines increases the need for a reliable assay system to evaluate their effectiveness. Previously, vaccine assays were carried out by individual manufacturers without access to standard reference preparations. A repository will allow study trends in the evolution of MDV to be studied.

Research Blood samples were collected from flocks in different parts of the country that have been experiencing MD losses. These samples were screened for the presence of serotype 1 MDV. Positive samples were stored in liquid nitrogen for future reference. In consultation with vaccine manufacturers and industry representatives, the Virology Laboratory set up a vaccine assay facility and is seeking accreditation with the National Association of Testing Authorities (NATA). During the establishment phase of the assay, a nominal cell count and virus titre, with maximum and minimum limits, were set for two reference preparations and were revised after their substantial use to monitor the stability of the assay. Studies were carried out in response to industry concerns about possible losses to vaccine potency from the widely used practice of using chilled diluent whilst administering vaccines. Another study assessed the differences between operators whilst performing vaccine assays.


Outcomes Some 300 blood samples were collected, of which 86 were positive for serotype 1

MDV. Characterisation studies did not commence within the project’s time frame. However, despite their pre-characterisation status, the samples remain as a viable repository of MDV isolates and will be available for comparative purposes in the future. A liquid overlay procedure was adopted for the vaccine assay and over 1000 assays have been conducted since the establishment of the facility in January 1998. Few manufacturers have used the facility despite its availability to all Australian manufacturers. An impending move to a new purpose-built facility and its audit, will take place before registration with NATA. The revised cell count and virus titre results indicated the stability of each reference preparation and thus validated the nominal virus titres used throughout the assay procedure. The use of diluent held at room temperature substantially decreases virus titre loss (as opposed to holding diluent at 4°C). Therefore, it is critical to maintain diluent at room temperature whilst administrating MD vaccines to maintain their potency. Differences in vaccine titre of parallel assays of two operators were non significant and, consequently, did not affect the assay result.

Publications Two presentations on the functions of the vaccine assay facility were given at meetings of the Australian Veterinary Poultry Association at Surfers’ Paradise in April 1999 and the Victorian Poultry Health and Welfare Liaison Group in September 1999.


Project Title

Candidate vaccine antigens against Pasteurella multocida

RIRDC Project No:

UMO-21A

Researcher: Prof Ben Adler Organisation: Monash University

Department of Microbiology Wellington Road CLAYTON VIC 3800


Objectives

• To evaluate the potential of genes of Pasteurella multocida as candidates for

attenuating mutations for the construction of live vaccines. • To assess the capacity of expressed recombinant antigens to protect against

infection in chickens. • To make recommendations with respect to vaccine choice and development

against fowl cholera.

Background

Fowl cholera caused by infection with the bacteria P. multocida is a cause of economic loss in the poultry industry in Australia and other countries. Available vaccines are undefined, empirically derived and of variable efficacy. There is a lack of knowledge of the basic mechanisms of immunity and pathogenesis (ability to cause disease) in P. multocida infections.

Research Several candidate antigens of P. multocida were expressed in recombinant form, purified and then tested for their ability to stimulate immunity against infection in the mouse model and in the natural chicken host. The outer-most component of the bacterial surface (termed the capsule) is critical in allowing bacteria to cause infection. The role of capsule in P. multocida infection and immunity was evaluated by the construction of defined acapsular mutants.

Outcomes Levels of immunity varied widely depending upon the antigen studied. The best prospect for a vaccine antigen appears to be the outer membrane protein Oma87. The capsule of P. multocida was shown to be a key virulence factor. Immunity could be stimulated by acapsular bacteria, indicating that recombinant vaccines based on protein antigens are feasible.

Implications The project has shown that it is possible to stimulate protective immunity against fowl cholera with either recombinant outer membrane proteins or with genetically attenuated bacteria. Further research is required for detailed evaluation of these approaches.

Publications Hunt, M.L., Boucher, D.J., Boyce, J.D. and Adler, B. (2001) In vivo expressed genes of Pasteurella multocida. Infect. Immun. In Press.

Chung, J.Y., Wilkie, I., Boyce, J.D., Townsend, K.M., Frost, A.J. and Adler, B. (2001) The role of capsule in the pathogenesis of fowl cholera caused by Pasteurella multocida serogroup A. Infect. Immun. In Press.


Boyce, J.D. and Adler, B. (2001) Acapsular Pasteurella multocida B:2 can stimulate protective immunity against pasteurellosis. Infect. Immun. 69, 1943-1946.

Townsend, K.M., Chung, J.Y., Boyce, J.D., Frost, A.J. and Adler, B. (2001) Genetic organisation of all Pasteurella multocida cap loci and its application in the development of a multiplex PCR typing system. J Clin Microbiol. In Press.

Hunt, M.L., Cox, A.J., Ruffolo, C.G., Rajakumar, K. and Adler, B. (2000) Characterisation of a Pasteurella multocida esterase gene which confers a haemolytic phenotype in Escherichia coli under anaerobic conditions. FEMS Microbiol. Lett. 192, 249-256.

Boyce, J.D. and Adler, B. (2000) The role of capsule in the pathogenesis of Pasteurella multocida B:2. Infect. Immun., 68, 3463-3468.

Boyce, J.D., Chung, J.Y. and Adler, B. (2000) Pasteurella multocida capsule: composition, function and genetics. J Biotechnol. 83, 153-160.

Boyce, J.D., Chung, J.Y. and Adler, B. (2000) Genetic organisation of the capsule biosynthetic locus of Pasteurella multocida M1404 (B:2). Vet Microbiol., 72, 121-134.

Mitchison, M., Wei, L., Kwang, J., Wilkie, I. and Adler, B. (2000) Overexpression and immunogenicity of the Oma87 outer membrane protein of Pasteurella multocida. Vet Microbiol., 72, 91-96.

Doughty, S.W., Ruffolo, C.G. and Adler, B. (2000) The type 4 fimbrial subunit gene of Pasteurella multocida, Vet Microbiol., 72, 79-90.

Cox, A., Ruffolo, C.G. and Adler, B. (2000) A Pasteurella multocida gene, ahpA, which confers haemolytic phenotype upon Escherichia coli. Vet Microbiol., 72, 135-152.

Hunt, M.L., Adler, B. and Townsend, K.M. (2000) The molecular biology of Pasteurella multocida. Vet Microbiol., 72, 3-25.

Adler, B., Bulach, D., Chung, J., Doughty, S., Hunt, M., Rajakumar, K., Serrano, M., van Zanden, A., Zhang, Y. and Ruffolo, C. (1999) Candidate vaccine antigens and genes in Pasteurella multocida. J Biotechnol., 73, 83-90.

Chung, J.Y., Zhang, Y., Adler, B. (1998) The capsule biosynthetic locus of Pasteurella multocida A:1. FEMS Microbiol. Lett., 166, 289-296.


Project Title

Effects of enzymes on gut microbial status in broiler chickens

RIRDC Project No:

UNE-70A

Researcher: Dr Mingan Choct Organisation: University of New England

School of Rural Science and Natural Resources - Animal Science ARMIDALE NSW 2351


Objective

• To identify opportunities for using commercial enzymes in broilers to modify

gut microbial status in a beneficial way to the host, in particular, in reduction of Clostridium perfringens.

Background

Broilers fed diets based on viscous grains such as wheat and barley appear to be more prone to necrotic enteritis than those fed non-viscous cereals such as corn and sorghum. It is hypothesised that the soluble non-starch polysaccharides (NSP) create a gut environment which encourages the proliferation of Clostridium perfringens, the causative agent for necrotic enteritis. Degradation of the NSP using enzymes thus may disrupt this environment, leading to reduced risk of necrotic enteritis outbreak.

Research A number of experiments was conducted to induce necrotic enteritis under experimental conditions and to examine the effect of xylanase supplementation on the gut microflora, including C. perfringens in broilers fed wheat based diets.

Outcomes Introduction of a low metabolisable energy (ME) wheat diet at the end of the starter phase of broilers led to a large increase in the number of C. perfringens. Xylanase supplementation markedly reduced the number of C. perfringens both in the ileum and the caeca. An experimental model system to induce necrotic enteritis required inoculation of both Eimeria spp and C. perfringens. The success rate of such a model was at best 60%.

Implications Change of diet at the end of the starter phase without antibiotics will leave the birds highly susceptible to an outbreak of necrotic enteritis. Use of appropriate enzymes may alleviate this problem should restrictions on the use of antibiotics in feed be implemented.


Project Title

Postgraduate scholarship – David Witcombe: Production and characterisation of recombinant antigens of Eimeria

RIRDC Project No:

UTS-3J

Researcher: Mr David Witcombe and Nicholas Smith Organisation: University of Technology

Molecular Parasitology Unit Dept of Cell and Molecular Biology PO Box 123 BROADWAY NSW 2007

Phone: (02) 9514 4013 Fax: (02) 9514 4026 Email : [email protected]

Objectives

• To prepare recombinant versions of a putative subunit vaccine candidate from

Eimeria, the 230 kDa merozoite protein. • To explore the potential molecular relationships between this protein and

gametocyte proteins of Eimeria. • To determine the importance of glycosylation of these proteins for induction of

immunity. • To determine how the genes encoding these antigens are organised and

expressed in the genome and whether they undergo antigenic variation or diversification.

• To assess whether combination immunoprophylaxis (eg vaccination with merozoite and gametocyte antigens) is more efficacious than vaccination with antigens from one developmental stage alone.

Background

Coccidiosis, caused by the protozoan parasite, Eimeria, is a major pathogenic disease of poultry worldwide. Coccidiosis costs the poultry industry internationally over $1 billion per year. The resistance of the parasite to anticoccidials, the high cost of development of new anticoccidials and the demands by the public for chemical-free meat drive the quest for a vaccine to control coccidiosis. Young chickens are protected against Eimeria by the transfer of antibodies from their mothers into the egg yolk, which is absorbed by hatchlings. These maternal antibodies have been used to identify proteins for inclusion in a vaccine. Exploration into the molecular composition of these proteins of Eimeria will allow deduction of the minimal components required for an effective vaccine. The development of a maternally-delivered vaccine against coccidiosis will benefit the poultry industry by allowing reduced use of chemotherapy, potentially effecting a significant cost reduction and satisfying consumer demand for residue-free meat.

Research The 230 kDa merozoite antigen of Eimeria maxima has been purified and N-terminal and internal tryptic digest amino-acid sequences determined. Using these sequences, the genetic code for the protein has been determined. Expression and production of recombinant proteins are underway. The sequences of gametocyte antigens (amino acid and genetic sequences) do not


appear to be related to the sequence of the 230kDa protein. There are several potential glycosylation sites in the 230 kDa merozoite protein. Similar sequences appear to be found in several species of Eimeria. Immunogenicity and efficacy trials using purified native and recombinant versions of the 230 kDa E. maxima merozoite protein will commence shortly.

Implications The results obtained to date suggest that the 230 kDa protein of E. maxima is a stage-specific molecule (found in asexual stages only). However, the observation that this protein is present in more than one species of Eimeria suggests that it has some conserved and important function in the development of the parasite. Therefore, it is a logical target for use in a subunit vaccine against coccidiosis.

Publications Witcombe, D., Belli, S., Wallach, M. and Smith, N. (2000) Western blot analyses and purification of an immunodominant asexual stage protein from Eimeria maxima. New Zealand and Australian Societies for Parasitology, Annual Scientific and General Meeting, Wellington, September.

Belli, S., Witcombe, D., Padula, M., Wallach, M. and Smith, N. (1999) The use of SDS-PAGE and 2-D PAGE in the analysis of parasite derived antigens. Australian Electrophoresis Society Sixth Annual Conference, Melbourne, October.

Witcombe, D., Belli, S., Wallach, M. and Smith, N. (1999) Characterisation of an immunodominant merozoite antigen from Eimeria maxima. Vaccines Against Animal Coccidioses, Interlaken, Switzerland, November.


Bird Nutrition and Feed Supply Project Title

Premium Grains for Livestock Program

RIRDC Project No:

GRD-1J

Researcher: Dr John Black Organisation: John L Black Consulting

Locked Bag 21 WARRIMOO NSW 2774


Objectives

• To identify the reasons for and magnitude of differences between grains in their

nutritional value for ruminants, pigs and poultry so that improvements in grain quality can be achieved by plant breeding and through grain processing and storage.

• To develop rapid tests, suitable for the site of grain receipt and/or use, to measure the nutritional value of grains so that they can be priced in accordance with their suitability as an animal feed.

• To develop a computer simulation model for ruminants to predict accurately the consequences of grain characteristics and of grain processing and storage on the productivity of feedlot cattle.

Background

The Program involved collaborative funding from the Grains, Pig and Dairy R&D Corporations, Meat and Livestock Australia and the Chicken Meat and Egg Programs of RIRDC. The Program is expected to improve the quality of grains available to the animal industries and to provide a more rational basis for trading feed grains based on measurement of the nutritional characteristics of grains determining their quality for different livestock enterprises.

Research Over 2000 grain samples covering the widest possible range in chemical and physical characteristics that may influence animal performance have been collected. The samples have been derived from germplasm collections, plant breeders, specially grown crops and commercial grains suspected of having extremes in nutritional values because of severe drought, frost damage or germination. All samples collected have been scanned with near infrared spectrometry (NIR). Approximately 115 analyses of chemical and physical characteristics thought to influence nutritional value have been conducted on all grains fed to animals. These involved analyses for individual carbohydrate, fatty acid and amino acid components, α- and β-amylase and anti-nutritional factors such as lectins, tannins and phytic acid. Physical properties included measurement of grain weight, hydration capacity, seed colour, seed diameter, seed size distribution, seed hardness index and profile, and the viscosity of whole grain, starch extract and acid soluble



extract. Light and electron microscopy has been used to examine the physical structure of some grains that differ markedly in their nutritional value. Many grains, including all those fed to animals have been examined using in vitro systems simulating both rumen fermentation and intestinal digestion. These analyses have been extremely useful for identifying grains that potentially have large differences in nutritional value for different classes of livestock and for screening potential processing procedures. A relatively small number of grains (approximately 100) covering the range identified in chemical and physical characteristics have been fed to sheep, cattle, pigs, broiler chickens and laying hens to determine the digestion of energy, individual grain compounds and, in some animal species, amino acids. The effects of processing and storage on the nutritional value of grain for different animal species have been evaluated using the in vitro systems. Development of rapid and accurate analytical tests for measuring the most important chemical and physical characteristics that determine nutritional value of feed grains has commenced. Preliminary analyses using NIR have been developed for predicting the digestible energy content of grains for pigs, apparent metabolisable energy (AME) for broiler chickens and whole animal dry-matter digestibility for sheep. In addition, NIR procedures have been used for predicting the major chemical components of grains and for predicting the in vitro digestion of various grain components.

Outcomes Results show that there is a wide variation in energy availability both within and between cereal grain species and between animal types. For example, the observed range in AME (MJ/kg) for broiler chickens for the grains examined in the Program is 15.4-16.1 for sorghum, 13.2-14.7 for wheat, 11.2-13.2 for barley, 11.2-14.4 for triticale, 12.6-13.4 for normal oat grain and 14.6 for naked oats. Naked oats had an AME value of 16.2 for laying hens. Sorghum has a much lower available energy content for cattle at 9.7 MJ/kg than for pigs (14.6 MJ/kg) or broiler chickens (15.9 MJ/kg). The energy value of waxy sorghum is enhanced considerable for cattle to 13.2 MJ/kg, but there was only a marginal increase of 0.1 to 0.5 MJ/kg for pigs and poultry.

Implications Several hypotheses about the factors determining the nutritional value of cereal grains for different animal types have been established and are being evaluated in the current Project GRD-3J.

Publications Balogun, R., Bird, S. H. and Rowe, J. B. (2000) Improving the nutritive value of sorghum grain by germination. Asian-Australian Journal of Animal Science. 13 (supp): 160.

Bird, S. H., Rowe, J. B., Choct, M., Stachiw, S., Tyler, P. and Thompson, R. D. (1999) In vitro fermentation of grain and enzymatic digestion of starch. Recent Advances in Animal Nutrition in Australia (Editor J. L. Corbett), 12: 53-61.

Black, J. L. (1999) Nutritional value of cereal grains for animals. Chemistry in Australia. 66: 7-9.

Black, J. L. (1999) The Premium Grains for Livestock Program. In: The Eleventh Australian Poultry & Feed Convention, Royal Pines Resort Gold Coast, Australia. pp. 226-32.

Black, J. L. (2001) Variation in nutritional value of cereal grains across livestock species. Australian Poultry Science Symposium, University of Sydney, Sydney, Australia 13: 22-9.


Choct, M., Hughes, R. J., Perez-Maldonado, R. and van Barneveld, R. J. (2001) The metabolisable energy value of sorghum and barley for broilers and layers. Australian Poultry Science Symposium, University of Sydney, Sydney, Australia 13: 39-42.

Dixon, R. M. and Stockdale, C. R. (1999) Associative effects between grains and forages: consequences for feed utilisation. Australian Journal of Agricultural Research 50: 757-73.

Evers, A. D., Blakeney, A. B. and O’Brien, L. (1999) Cereal structure and composition. Australian Journal of Agricultural Research. 50: 629-50.

Farrell. D. J. (1999) In vivo and in vitro techniques for the assessment of the energy content of feed grains for poultry: a review. Australian Journal of Agricultural Research 50: 881-8.

Flinn, P. C. (2000) Current and potential use of NIR in the fodder and grain industries: a ruminant’s perspective. In: Reading more from the spectrum, 9th Australian Near Infrared Spectroscopy Conference, Horsham, VIC, Apr 2000.

Flinn, P. C. (2000) Rapid methods for the quality assessment of grains for animal nutrition. In: Chemical aspects of grain in human and animal nutrition., Cereal Chemistry Division Symposium, 11th RACI Convention, Canberra, ACT, Feb 2000, p15.

Flinn, P. C., Heazlewood, P. G. and Dalton, S. L. (2000) Recent developments in improving the prediction of digestibility of feed grains. In: 9th International Conference on Near Infrared Spectroscopy Verona, Italy, Jun 1999 (in press).

Hogan, J. P. and Flinn, P. C. (1999) An assessment by in vivo methods of grain quality for ruminants. Australian Journal of Agricultural Research 50: 843-54.

Hughes, R. J. and Choct, M. (1999) Chemical and physical characteristics of grains related to variability in energy and amino acid availability in poultry. Australian Journal of Agricultural Research 50: 689-701.

Hughes, R. J., Choct, M. and van Barneveld, R. J. (2001) Factors influencing the energy values of Australian cereal grains fed to broilers. Australian Poultry Science Symposium, University of Sydney, Sydney, Australia 13: 30-8.

Kaiser, A. G. (1999) Increasing the utilisation of grain when fed whole to ruminants. Australian Journal of Agricultural Research. 50: 737-56.

Kitessa, S., Flinn, P. C. and Irish, G. G. (1999) Comparison of methods used to predict the in vivo digestibility of feeds in ruminants: A review. Australian Journal of Agricultural Research 50: 825-41.

Kruk, J. A. and van Barneveld, R. J. (1999) Near infra-red spectrophotometry predictions of digestible energy in cereals for pigs. In: Manipulating Pig Production VII (submitted, Editor P. D. Cranwell) Australasian Pig Science Association, Werribee, VIC.

Nagorka, B. N. (2000) A ruminant model to estimate the nutritive value of grains in cattle feedlots. International Report, CSIRO CLI.

Nagorka, B. N., Gordon, G. L. R. and Dynes, R. A. (2000) Towards a more accurate representation of fermentation in mathematical models of the rumen. Modelling Nutrient Utilization in Farm Animals (Editors McNamara, J. P., France, J. and Beever, D. E.) CAB International, London (in press).

Moughan, P. J. (1999) In vitro techniques for the assessment of the nutritive value of feed grains for pigs: a review. Australian Journal of Agricultural Research 50: 871-9.

O’Brien, L. (1999) Genotype and environment effects on feed grain quality. Australian Journal of Agricultural Research. 50: 703-19.

O’ Brien, L. (2000) Genotype and environment effects on feed grain quality. Royal Chemical Institute, 11th National Convention, Canberra, ACT, Feb 2000.

O’Brien, L. and Blakeney, A. B. (2000) Genetic variation for components of dietary fibre in wheat and barley grains. 11th Cereals and Bread Congress, Gold Coast, Sept 2000.


O’Brien, L. and Blakeney, A. B. (2000) Opportunities for genetically improving the dietary fibre content of wheat and barley. International Dietary Fibre Congress, Dublin, Ireland, May 2000.

O’Brien, L., Tredea, A. M., van Barneveld, R., Bird, S. and Rowe, J. (2000) Genotype and environment effects on measures of feed grain quality in wheat, barley and oats. 11th Cereals and Bread Congress, Gold Coast, Sept 2000.

Petterson, D. S., Harris, D. J., Rayner, C. J., Blakeney, A. B. and Choct, M. (1999) Methods for the analysis of premium livestock grains. Australian Journal of Agricultural Research 50: 775-87.

Ravindran, V. and Bryden, W. L. (1999) Amino acid availability in poultry – in vitro and in vivo measurements. Australian Journal of Agricultural Research 50: 889-908.

Rowe, J. B., Choct, M. and Pethick, D. W. (1999) Processing cereal grains for animal feeding. Australian Journal of Agricultural Research 50: 721-36.

van Barneveld, R. J. (1999). Chemical and physical characteristics of grains related to variability in energy and amino acid availability in pigs: a review. Australian Journal of Agricultural Research 50: 667-87.

van Barneveld, R. J. (1999) Controlling the nutritional quality of stockfeed ingredients. VICTAM Feed Ingredients and Grain Processing Asia ’99. Bangkok, Thailand, 3-5 November, 1999.

van Barneveld, R. J. (1999) Physical and chemical contaminants in grains used in livestock feeds. Australian Journal of Agricultural Research 50: 807-23.

van Barneveld, R. J. (1999) Strategies for the assessment of livestock feed ingredient quality. Recent Advances in Animal Nutrition in Australia 12: 63-72.

van Barneveld, R. J., Ru, Y. J., Wyatt, G. F. and Pluske, J. R. (2000) Relationship between the ileal and faecal digestible energy content of pig diets containing Australian barley cultivars. Proceedings of the Nutrition Society of Australia (in press).

van Barneveld, R. J., Ru, Y. J., Szarvas, S. R. and Wyatt, G. F. (1999) Range in digestible energy and true ileal digestible lysine content of 11 barley samples. In: Manipulating Pig Production VII (submitted, Editor P. D. Cranwell) Australasian Pig Science Association, Werribee, VIC.

van Barneveld, S. (1999) Chemical and physical characteristics of grains related to variability in energy and amino acid availability in ruminants: a review. Australian Journal of Agricultural Research 50: 651-66.

White, C.L. and Ashes, J. R. (1999) A review of methods for assessing the protein value of grain fed to ruminants. Australian Journal of Agricultural Research 50: 855-69.

Wrigley, C. M. (1999) Potential methodologies and strategies for the rapid assessment of feed-grain quality. Australian Journal of Agricultural Research 50: 789-805.

Zarrinkalam, M. R., van Barneveld, R. J., Tivey, D. R. and Choct, M. (1999 Predicting energy availability in barley for pigs and poultry using rapidly determined fibre content. In: Manipulating Pig Production VII (submitted, Editor P. D. Cranwell) Australasian Pig Science Association, Werribee, Vic.


Project Title

Postgraduate scholarship- Andreas Kocher: Increasing the nutritive value of grain legumes for poultry by use of more efficacious enzyme systems

RIRDC Project No:

UNE-64A

Researcher: Mr Andreas Kocher and Dr Mingan Choct Organisation: Universtiy of New England

School of Rural Sciences ARMIDALE NSW 2350


Objective

• To investigate the effect of commercial feed enzymes on the nutritive value of

grain legumes.

Background

Grain legumes and oilseed meals are already widely used as the main protein source in pig and poultry diets. However, the high content of indigestible carbohydrates such as oligosaccharides and non-starch polysaccharides (NSP) in these ingredients will reduce their nutritive value for broiler chickens. In diets based on wheat and barley, high levels of soluble NSP raise digesta viscosity in the intestine of chickens leading to reduced starch, protein and lipid digestion. The addition of commercial feed pentosanases and β-glucanases to these diets generally results in a significant reduction in intestinal viscosity and subsequently in an increase in growth performance. There is only limited documentation available on the anti-nutritive effects of NSP in legumes and oilseed meals and the possible benefits from the addition of commercial feed enzymes. The objectives of the work reported in this PhD project were to investigate the effects of commercial feed enzymes on the nutritive value of vegetable proteins. In particular, this study has focused on the effects of feed enzymes on the utilisation of neutral non-starch polysaccharides of lupins, soybean, canola and sunflower seeds.

Research A preliminary study investigated the effects of feed enzymes designed to hydrolyse NSP in vegetable proteins on the utilisation of NSP in a low NSP cereal (sorghum). In four separate studies the effects of commercial enzyme products on the nutritive value of lupins (L. angustifolius and L. albus), canola meal, sunflower meal and soyabean meal were investigated. These studies were designed to determine the effects of enzyme addition on the performance and nutrient utilisation of broilers, as well as on the composition of NSP in the jejunum and ileum of broilers fed diets containing a minimum of 35% legumes with or without enzyme supplementations.

Outcomes The addition of commercial feed enzymes designed to utilise NSP of legumes had no effects on broiler performance, apparent metabolisable energy (AME) and the composition of NSP in the intestine of broilers fed a sorghum/casein basal diet. It was concluded that, in future experiments, any differences between diets with or without enzymes would be entirely due to the vegetable protein component. In diets with L. Angustifolius the addition of enzymes with a high level of



polygalacturonase activity significantly (P<0.001) raised digesta viscosity and increased the concentration of soluble NSP in all sections of the intestine. The structures of NSP isolates from ileal digesta were characterised using size-exclusion chromatography and 1H-NMR spectroscopy. The structural composition of isolates obtained from diets with or without enzyme supplementation revealed that the enzyme solubilised parts of the pectic backbone, most likely the main pectic polymer, rhamnogalacturonan in lupins. The actual cleavage point of the enzyme was found to be in the less branched regions of the pectic backbone. Enzymes tended to reduce the water-soluble NSP fraction in the upper intestine of broilers fed diets high in canola meal, sunflower meal and soya bean meal. However, only the addition of these enzymes at a very high dosage (five times the recommended dosage) led to a significant (p<0.05) improvement in AME and nutrient digestibility. Benefits of enzyme addition to commercially formulated broiler diets with high levels of canola meal were found in improved drip loss, dress yield and breast meat.

Implications Commercial feed enzymes with increased polygalacturonase activities have noticeable effects on the carbohydrate components of the vegetable proteins. However, actual commercial benefits were only evident when enzymes were added at very high dosage level (five times the recommended dosage). The results of this project demonstrated that soluble NSP of vegetable proteins do not exhibit strong anti-nutritive effects and can be largely utilised throughout the gut by microbial digestion. The benefit of feed enzyme addition lies in the increase in efficiency of using NSP as energy sources in broiler diets. The depolymerisation of pectins by exogenous enzymes in the upper intestine will give the bird access to intracellular entrapped nutrients as well as provide a more efficient energy utilisation.

Publications Kocher, A., Hughes, R.J., Choct, M. and Broz, J. (2000) Effect of food enzyme addition on utilisation of lupin carbohydrates by broilers. British Poultry Science, 41: 75-82.

Kocher, A. and Choct, M. (2000) Enzyme supplementation of diets containing high levels of legumes. Proceedings of the 12th Australian Poultry Science Symposium, 12: 159-163.

Kocher, A. and Choct, M. (2000) Lupin non-starch polysaccharides: Utilisation by chickens in the presence of feed enzymes. Dietary Fibre 2000, Dublin Ireland, p. 105.

Choct, M. and Kocher, A. (2000) Use of enzymes in non-cereal grain feedstuff. Proceedings of the World Poultry Science Congress, 11: S1.10.04.

Choct, M. and Kocher, A. (2000) Non-starch polysaccharides: Digestion and its secondary effects in monogastrics. Proceedings of the 24th annual scientific meeting of the Nutrition Society of Australia, 24: 31-38.

Kocher, A., Choct M., Porter, M.D. and Broz, J. (2000) The effects of enzyme addition to broiler diets containing high levels of canola and sunflower meal. Poultry Science, 79: 1767-1774.

Kocher, A. and Choct, M. (2001) Do enzymes improve the nutritive value of soybeans in broiler diets? Proceedings of the 13th Australian Poultry Science Symposium 13:204-207.

Kocher, A., Choct, M., Morrisroe, L. and Broz, J. (2001) Effects of enzyme supplementation on the replacement value of canola meal for soybean meal in broiler diets. Australian Journal of Agricultural Research, 54: 447-452.


Kocher, A., Choct, M., Porter, M.D. and Broz, J. (2001) Effect of food enzyme addition on utilisation of soybean carbohydrates by broiler. British Poultry Science, (in press).

Kocher, A. (2001) Enzymatic Degradation of Non-Starch Polysaccharides in Vegetable Proteins in Poultry Diets. Recent Advances in Animal Nutrition in Australia ‘2001 (in press).


Project Title

Postgraduate scholarship – Ron Newman: Manipulation of lean tissue deposition in broiler chickens by altering the sensitivity of tissues to insulin

RIRDC Project No:

US-68A

Researcher: Mr Ron Newman and A/Prof Wayne L Bryden Organisation: University of Sydney

Department of Animal Science Werombi Road CAMDEN NSW 2570


Objective

• To investigate strategies, based on altering the fatty acid composition of the diet,

for reducing fat deposition and increasing lean meat deposition in broilers.

Background

The selection for rapid growth of the modern broiler strains has been accompanied by increased fat deposition. Glucose-insulin balance has been suggested as the main reason for the increased adiposity in broilers. It has been shown that dietary fatty acid profile can alter adiposity in mammals by changing tissue insulin sensitivity and this was studied in birds in the project.

Research Studies were designed to examine the effects of n-3 (fish oil) and n-6 (sunflower oil) polyunsaturated fatty acids in comparison to a saturated fat (tallow) on physiological and morphological characteristics of broilers.

Outcomes Research undertaken showed that broilers fed either of the polyunsaturated fats (fish or sunflower oils) had a significant reduction in fat pad mass and an increase in breast muscle weight compared to broilers fed tallow. The changes in body composition were without penalty on weight gain and feed conversion efficiency. The n-3 and n-6 fats achieved reduced lipid deposition through different mechanisms.

Implications The project has shown that simple dietary manipulation can alter carcass composition and potentially have a positive impact on the economics of broiler production, especially the further processing trade. Further studies are required to determine the optimal fatty acid or combination of fatty acids that reduce lipid deposition.

Publications Newman, R.E., Downing, J.A., Dehon, J.A. and Bryden, W.L. (1998) Manipulation of glucose metabolism in the broiler chicken with dietary fatty acids. Proc. Aust. Poult. Sci. Symp. 10: 210.

Newman, R.E., Downing, J.A., Bryden, W.L., Fleck, E., Buttemer, W.A. and Storlien, L.H. (1998) Dietary polyunsaturated fatty acids of the n-3 and the n-6 series reduce abdominal fat in the chicken (Gallus domesticus). Proc. Nutr. Soc. Aust. 22: 54.


Newman, R.E., Downing, J.A., Jackson, C.M. and Bryden, W.L. (1999) Differences in glucose metabolism between broiler and layer chickens. Proc. Aust. Poult. Sci. Symp. 11: 164.

Newman, R.E., Fleck, E., Downing, J.A., Ashes, J.R., Storlien, L.H. and Bryden, W.L. (1999) Dietary n-3 polyunsaturated fatty acids alter avian glucose metabolism. Proc. Nutr. Soc. Aust. 23: 62.

Bryden, W.L., Newman, R.E. and Downing, J.A. (1999) The changing role of fat in poultry nutrition. 4th Technical Symposia, Novus International, Leura, NSW, pp.10.

Newman, R.E. (1999) Manipulation of growth in animals and poultry. 40th Anniversary Seminar, Poultry Research Foundation, University of Sydney, p. 9.

Newman, R.E., Downing, J.A., Storlien, L.H., Fleck, E., Ashes, J.A. and Bryden, W.L. (2000) Dietary n-3 and n-6 fatty acids alter pituitary sensitivity to GHRH infusion but not to LFRH infusion in the domestic chicken (Gallus domesticus). Proc. AAAP-ASAP 2000 Congress, Sydney. (In press).

Newman, R.E., Downing, J.A., Storlien, L.H., Fleck, E., Ashes, J.A. and Bryden, W.L. (2000) Dietary n-3 and n-6 fatty acids alter pituitary sensitivity to GHRH infustion but not to LHRH infusion in the domestic chicken (Gallus domesticus). Asian-Aust. J. Anim. Sci. (Supplement A) 13: 213.


Food Safety Project Title

Risk factors for Campylobacter spp. in Australian broilers

RIRDC Project No:

DAQ-245A

Researcher: Ms Jeanette Miflin Organisation: Department of Primary Industries (Qld)

Animal Research Institute Locked Mail Bag No 4 MOOROOKA QLD 4105


Objective

• To reduce the levels of Campylobacter spp. in Australian broilers by the

development of on-farm HACCP plans that are based on risk factors relevant to Australian conditions.

Background

Campylobacter spp. are a major human health concern, as they are the leading cause of food-borne gastroenteritis in Australia. Raw or undercooked poultry meat is considered to be an important source of human infection. In order to control the level of contamination of poultry products, the industry needs to understand when and how campylobacters gain access to poultry flocks.

Research A survey of 56 broiler farms was conducted to determine the prevalence of Campylobacter spp. prior to partial depopulation. Longitudinal studies were then undertaken to address questions relating to source and spread of Campylobacter colonisation within broiler flocks. Study farms were sampled weekly or more frequently from placement until final slaughter. Samples were collected from chickens (23,465) as well as from potential sources inside and outside the shed. Isolates of C. jejuni and C. coli were subjected to a DNA typing procedure (flaA typing).

Outcomes More than 50% of farms were Campylobacter-free until partial depopulation. When farms became colonised before partial depopulation, first positive samples were only detected when the chickens were 24 days or older. The within-shed rate of spread was rapid, with the number of positive samples increasing from 1/100 to 99/100 within three days. Vertical transmission from broiler breeders to day-old chicks did not occur. Within-shed sources such as drinking water and darkling beetles were not the primary source of introduction to the flock, although they were important agents of spread within the flock. Litter re-use from batch to batch may have played a role in carryover of the organism in some instances. Multiple sources of the organism external to the shed were identified. Waterbirds,


mice and domestic animals (cattle and sheep) were shown to carry C. jejuni. The use of flaA typing provided conclusive proof that crates used at partial depopulation introduced the organism to previously negative flocks. Farms with good biosecurity can maintain Campylobacter-free status at least to partial depopulation. During the course of the study, three farms remained Campylobacter-free to final slaughter. The project demonstrated that good biosecurity (in particular the use of properly maintained footbaths), and good crate cleaning procedures, can prevent C. jejuni colonisation of broiler flocks under Australian production conditions.

Implications It is possible for Australian growers to produce Campylobacter-free chickens provided that adequate on-farm biosecurity is maintained and that pick-up crates are free of contamination.

Publications Miflin, J.K. (1999) Prevalence of, and risk factors for, Campylobacter in meat chickens. Food Safety Risk Analysis Workshop for Primary Industries, Canberra.

Miflin, J.K, Blackall, P.J. and More, S.J. (1999) Preliminary epidemiological studies on Campylobacter spp. in meat chickens in Queensland, Australia. 10th International Workshop on Campylobacter, Helicobacter and related organisms, Baltimore, USA.

Miflin, J.K. and More, S.J. (2000) Campylobacter and the meat chicken industry. Poultry Information Exchange, Gold Coast, Queensland.

Miflin, J.K., Templeton, J.M. and More, S.J. (2000) The epidemiology of Campylobacter spp. in poultry. Scientific Meeting, Australian Veterinary Poultry Association, Gold Coast, Queensland.

Miflin, J.K. and More, S.J. (2000) Campylobacters, poultry and people. Annual Scientific Meeting, Australian Society for Microbiology, Cairns, Queensland.

Miflin, J.K., Templeton, J.M. and More, S.J. (2000) Longitudinal studies of Campylobacter spp. on broiler farms in South East Queensland. Queensland Poultry Science Symposium, Gatton, Queensland.

Miflin, J.K., Templeton, J.M. and More, S.J. (2001) Epidemiological studies of Campylobacter colonisation of broiler flocks in South East Queensland. Australian Poultry Science Symposium, Sydney.


Project Title

Molecular basis of benign colonisation of Salmonella Sofia in chickens

RIRDC Project No:

IMVS-1A

Researcher: Dr Michael Heuzenroeder, Mr Christopher Murray, Ms Rina Dalcin and Dr Mary Barton

Organisation: Institute of Medical and Veterinary Science Infectious Diseases Laboratories Box 14 Rundle Mall P.O. ADELAIDE SA 5000


Objectives

• To continue to offer to industry the conventional and molecular typing service

developed in the course of previous RIRDC projects. • To evaluate Salmonella Sofia strains that have been characterised genetically

(ie. rationally chosen) for their ability to colonise and competitively exclude virulent serovars eg. Typhimurium.

• To identify the bacterial factors involved in the efficient colonisation of chickens by S Sofia and to genetically characterise them by DNA sequence analysis.

Background

Contamination of poultry products by Salmonella can cause public health problems. Because of this challenge, the development of rapid typing methods for Salmonella in conjunction with classical methods was initiated. Molecular methods offer greater discriminating power between human and poultry isolates for epidemiological studies of disease transmission. It had also been observed that Salmonella Sofia has become the major isolate from chickens, but is rarely seen in humans. This suggests that this organism is non disease causing and could be naturally excluding disease causing Salmonella from chickens.

Research Conventional and molecular typing methods (pulsed field gel electrophoresis or PFGE) were used to process relevant isolates from industry sources for monitoring of Salmonella in chicken flocks. S. Sofia strains able to colonise chickens for extended periods (up to 36 days) were identified. It was shown that S. Sofia strain MH76 could co-colonise chickens with S. Typhimurium but not exclude it. The mechanism of colonisation of chickens by S. Sofia is most probably determined by factors on the bacterial cell surface. DNA sequences potentially encoding genes for colonisation factors may explain the ability of S. Sofia to colonise chickens.

Outcomes Other than for the observation that S. Virchow is being found with increasing frequency in the southern states, the distribution of other serovars remains largely unchanged. S. Sofia does not appear as a competitive exclusion agent for some pathogenic Salmonella. S. Sofia strain MH76 has been identified as a strain that could be a potential carrier for antibacterial products. Novel genes for colonisation factors in S. Sofia have also been identified.

Implications Epidemiological and scientific evidence continues to support the notion that S. Sofia,


which remains the most common Salmonella colonising chickens used for chicken meat production, is of low virulence to humans. The discovery of a gene not predicted to be present in a S. Sofia is very interesting. These genes encode factors implicated in the invasion of the lymphoid tissue in the gut by virulent Salmonella. It could be hypothesised that the presence of these genes may partially explain the continued colonisation of Australian chickens with S. Sofia.

Publications Heuzenroeder, M.W. (1998) Salmonella Sofia Colonisation Factors. Australian Veterinary Poultry Association Seminar Series. Surfers Paradise, 22-23rd April 1998.

Heuzenroeder M.W. et al. (1998) Molecular typing of Salmonella. Is there a phage in the ointment? Proceedings of the Fourth Asia Pacific Poultry Health Conference, page 144 Melbourne 1998

Heuzenroeder, M.W. (2000) Beneficial Salmonella Sofia colonisation of Australian chickens? Society for Applied Microbiology News 1:32 March 2000



Project Title

Use of avirulent Campylobacter jejuni strains to control poultry-derived campylobacter food poisoning

RIRDC Project No:

RMI-7A

Researcher: Dr V Korolik* and Professor PJ Coloe Organisation: RMIT University

Department of Applied Biology and Biotechnology GPO Box 2476V MELBOURNE VIC 3001 *Griffith University School of Health Science PMB 50 GOLD COAST MAIL CENTRE QLD 9726

Phone: (07) 5594 8321 Fax: (07) 5594 8908 Email: [email protected] Internet: http://www.gu.edu.au/home/home.html

Objectives

• To detect and identify those strains of Campylobacter jejuni that colonise

chickens but do not constitute a disease problem in humans. • To identify or construct a selection of non-virulent strains which are able to

colonise chickens, but not cause disease and to fully characterise those strains and test them for their ability to exclude other, potentially virulent strains.

Background

Campylobacter species are now well recognised as one of the major causes of enteric disease in humans. Indeed Campylobacter species, in particular C jejuni, are now the most common cause of food borne disease in the developed world and have surpassed Salmonella and Shigella spp as causes of lost production in the workplace. Campylobacter disease is zoonotic since the organisms are widespread in animals and birds where, by and large, they are commensals. However, the processing of poultry and other animal food products often leads to contamination of the end product and consumption of undercooked meats leads to the transfer of campylobacter disease to humans

Research Previous work demonstrated that there are differences between strains of C.jejuni in their ability to colonise chickens. Some strains do not colonise chickens while others not only colonise, but are able to displace resident strains. One such strain was shown to be a very strong coloniser of the chicken intestine, could displace other, well established colonising campylobacters and also once established in the chicken intestinal tract, this strain could not be displaced by other strains. A chick colonisation model that can be used to determine the ability of isolates to colonise and to persist in chickens and to displace other isolates had also been developed. A range of Campylobacter strains from both humans and chickens was tested for their ability to invade human intestinal cells in tissue culture and established a set of characterised invasive and a set of non invasive strains. A DNA based Polymerase Chain Reaction (PCR) test that can detect minute


numbers of Campylobacter bacteria in faeces and on poultry destined for human consumption was developed. The assay is quick and accurate and can be applied at any stage in the production and marketing stage.

Outcomes Molecular identification tests were developed to detect, identify and differentiate C.jejuni strains from chicken and human sources. Chicken colonisation models have been established and a super-colonising C.jejuni strain was assessed. Establishment of characterised sets of Campylobacter strains for colonisation and invasion. Isolation of C.jejuni virulence determinants and genes encoding invasion.

Implications An alternative approach to total exclusion of normal flora campylobacters from chickens is to use an avirulent strain of C.jejuni, which has been fully characterised and which presents no danger to humans, to establish continuing infection in chickens that does no present danger to humans. A PCR detection test for C.jejuni was developed that could be used to quickly and accurately to screen human faecal samples in the hospital pathology labs and chicken flocks for presence of Campylobacter spp, differentiation of C.jejuni from other campylobacters and to determine to which polymorphic group the strains belong.

Publications Korolik, V., Fry, B.N., Alderton, M.R, van der Zeijst, B.A.M. and Coloe P.J. (1997) The expression of Campylobacter hyoilei lipooligosaccharide (LOS) antigens in Escherichia coli. Microbiology, 143: 3481-3489.

Korolik, V., and Coloe, P.J. (1997) Tummy ache bugs. 1997. Microbiology Australia, 18:16-17.

Korolik, V., Alderton, M.R., Chang, J., Smith, S.S. and Coloe, P.J. (1998) Isolation and molecular analysis of colonising and Non-colonising strains of Campylobacter jejuni and Campylobacter coli following experimental infection of young chickens. Veterinary Microbiology, 60: 239-249.

Fry, B.N., Korolik, V., Pennings, M.T., ten Brinke, J.A., Coloe, P.J., and van der Zeijst, B.A.M. (1998) Identification and characterization of the Lipopolysaccharide biosynthesis locus of Campylobacter jejuni 81116. Microbiology 144: 2049-2061.

Korolik, V., Chang, J. and Coloe, P.J. (1998) Differences in colonisation potential of Campylobacter jejuni strains in chickens In: Campylobacter IX. Ed: Lasovica, AJ. and Newell, D.G. 365-368.

Fry, B.N., ten Brinke, J.A., Korolik, V. and van der Zeijst, B. (1998) The lipopolysaccharide biosynthesis locus of Campylobacter jejuni 81116. In: Campylobacter IX. Ed: Lasovica, AJ. and Newell, D.G. :503-508.

Lee, A., Smith, S.C., Michalski, W., Shiells, B., Korolik, V. and Coloe, P.J. (1998) Purification and characterisation of a novel Campylobacter jejuni cytotoxin. In: Campylobacter IX. Ed: Lasovica, AJ. and Newell, D.G. : 300-303.

Clow, K., Park, S.F., Hawtin, P.R., Korolik, V. and Newell D.G. (1998) The genotypic and phenotypic comparison of Campylobacter jejuni strains from poultry and humans. In: Campylobacter IX. Ed: Lasovica, AJ. and Newell, D.G. : 368-369.

Fry B.N., Yuen-Yuen Chen, S.F., Newell, D.G., Coloe, P.J. and Korolik, V. (2000) The galE gene of Campylobacter jejuni is involved in LPS synthesis and virulence. Infection and Immunity, 68: 2594-26091.

Fry, B.N., Oldfield, N.J., Korolik, V., Coloe, P.J. and Ketley, J.M. (2000) The genetics of lipopolysaccharide biosynthesis of Campylobacter. In: Campylobacter 2nd Edition. Eds: I. Nachamkin and M.J. Blaser. 381-403. ISBN: 1-55581-165-5.


Smith, M.A., Finel, M., Korolik, V. and Mendz G. L. (2000) Characteristics of the NDH-1 complex and terminal oxidases of the microaerophilic bacteria C. jejuni and H. pylori. Archives of Biochemistry, In press.

Dep, M.S., Mendz , G.L., Smith, M.A., Coloe, P.J., Fry, B., and Korolik, V. (2000) Differentiation between Campylobacter hyoilei and Campylobacter coli using genotypic and phenotypic analyses. Accepted to Journal of Systematic and Environmental Microbiology.


Project Title

Antibiotic resistance in bacteria isolated from poultry

RIRDC Project No:

USA-9A

Researcher: A/Prof Mary Barton Organisation: University of South Australia

School of Pharmacy and Medical Sciences GPO Box 2471 ADELAIDE SA 5001


Objective

• To assess the resistance to antibiotics of bacteria from chicken carcasses and to

investigate the antibiotic resistance patterns of Clostridium perfringens, the cause of necrotic enteritis in chickens.

Background

Antibiotic resistance in bacteria that can potentially cause disease in humans is of major concern world-wide. Although most resistance relates to misuse and overuse of antibiotics in hospitals and the community generally, there is concern about transfer of antibiotic resistant bacteria from animals to humans via the food chain or direct contact. Some antibiotics may be withdrawn from use in animals because of concerns about resistance and this presents some problems in the control of necrotic enteritis in particular.

Research Strains of salmonella, Escherichia coli, campylobacter and enterococci and Clostridium perfringens were isolated from chicken carcass rinse samples and chicken intestines. They were assessed for resistance against a range of antibiotics used to treat and prevent infections in chickens and people and against some growth promotants and coccidiostats with antibacterial activity, including those of concern to medical authorities. Carcass rinse samples came from two company laboratories and intestinal samples from a company processing plant. Salmonella isolates were also obtained from a reference laboratory.

Outcomes Antibiotic resistance was found to be quite widespread in all the isolates tested. Overall, resistance patterns were not dissimilar to those reported overseas but detailed comparisons are of limited value because of the differences in antibiotic use regimes. No fluoroquinolone resistance was detected and, although not surprising as no fluoroquinolones are registered for use in poultry or livestock, the finding was pleasing because this is a key human antibiotic. Vancomycin resistant enterococci (VRE) of the type predominant in Europe was found – again not unexpected as avoparcin which selects for this type of resistance has been used in meat chickens in Australia. The type of VRE which causes the majority of human infections in Australia was not detected. Resistance to virginiamycin, which is closely related to dalfopristin/quinupristin (a human antibiotic), was also detected. Clostridium perfringens isolates were resistant to most of the growth promotant and coccidiostat antibiotics tested.

Implications The chicken industry needs to continue to promote responsible use of antibiotics by companies and growers. Resistant strains of salmonella and campylobacter which


are recognised causes of food poisoning are present in Australian chickens and, although antibiotics are rarely used to treat food poisoning, it is not a desirable situation for the industry. VRE are also present but numbers should decline as avoparcin is no longer available in Australia. Virginiamycin resistance in enterococci is a potential problem and could lend weight to the arguments of those who would like to see it banned in animals. Antibiotic treatment is clearly not a long-term option for control of necrotic enteritis as C. perfringens strains in this study have already acquired resistance to most of those available. Salinomycin, narasin and avilamycin may be useful in the short to medium term.


Animal Welfare

Project Title

A welfare audit for the transport and processor sectors of the broiler industry

RIRDC Project No:

DAV-162A

Researcher: Dr John Barnett Organisation: Department of Natural Resources & Environment (Vic)

Agriculture Victoria VIAS (Animal Welfare Centre) Private Bag 7, Sneydes Road WERRIBEE VIC 3030


Objectives

• To provide audit documentation for the pick-up, transport and processor sectors

of the chicken meat industry and thereby complete audit documentation for all sectors of the chicken meat industry.

• To finalise draft audit documentation from previous project DAV-147A on the hatchery, broiler, breeding and pullet rearing sectors of the chicken meat industry, following industry consultation and feedback.

• To evaluate audit documentation already completed in project DAV-147A at commercial farms.

Background

Quality Assurance programs within the chicken meat industry predominantly focus on animal health and food safety and there was a need, coinciding with a more informed and demanding customer base, to expand these programs to include animal welfare issues so that industry remains sustainable into the next century. A large number of the issues that producers/unit managers focus on daily, including animal health, production and food safety, also relate to animal welfare, although this information was not previously formalised within one document. Audit documentation provides some certainty for all staff involved, so that they, the company, consulting veterinarians, the public and any internal or external auditors are asking the same questions in terms of animal care. Broiler companies already provide considerable information on maintaining high levels of animal welfare and producers, in their daily tasks, already largely implement this information. One purpose of this project was to put all this information together and add additional information from the literature and experts in Australia. This project on the pick-up, transport and processing sectors of the industry completes the industry audit documentation. This audit for the Australian chicken meat industry is the first comprehensive welfare audit for an industry that has been developed to cover all sectors.

Research The details of the audit documentation were directed and determined by a management group including researchers and people with technical expertise in



broiler production, welfare issues in the industry, general community views on welfare issues and legal requirements. The management group developed a set of audit questions and supporting material relating to welfare in the commercial environment that could be completed by a yes/no answer with appropriate documentation for the required record keeping. The documentation was evaluated in a study at 12 treatment and 12 control broiler farms for three batches of birds.

Outcomes The documentation for auditing welfare for the chicken meat industry is complete. It covers all sectors of the industry: hatchery, broilers, broiler rearers, broiler layers and pick-up, transport and processing. It is available from RIRDC as a hard copy and on CD-rom. The evaluation trial showed improved record keeping and lower mortality in the first week after placement at the treatment farms, indicating the benefits of implementing the audit process.

Implications There are some short term and long term benefits of the project. The short term benefits are those to be gained from advertising industry’s proactive position on animal welfare to the community generally. The long term benefits will only be achieved by implementation of the audit process. These benefits will include an ability by industry to demonstrate compliance with relevant Codes of Practice and many targets that are of a higher standard than the Codes of Practice, improvements in compliance levels with targets over time, an ability to identify and solve problems on individual farms and where necessary initiate industry education on specific issues and by these actions a reassurance for the public on welfare standards within the chicken meat industry.

Publications Barnett, J.L. (1999) A welfare audit for the broiler industry. Final report to the Rural Industry Research and Development Corporation, project number 147A.

Barnett, J., Glatz, P. and Almond, A. (1999) A welfare audit for the grower, hatchery and laying sectors of the chicken meat industry in South Eastern Australia. Proceedings South Australian Pig and Poultry Fair,Roseworthy, p. 44.

Barnett, J., Glatz, P. and Almond, A. (1999) A welfare audit for the transport and processing sectors of the chicken meat industry – research in progress. Proceedings South Australian Pig and Poultry Fair, Roseworthy, p. 45.

Barnett, J.L. (2000) A welfare audit for the broiler industry: A model for other animal industries. Agriculture Victoria Institute Group Research Conference, Poster 1-21.

Barnett, J.L. (2000) Practical lessons learned from the development of Chicken Meat QA. Layer Hen Housing Conference (July, Gold Coast, Qld), (AFFA, Canberra), Attachment N.

Barnett, J.L., Almond, A. and Glatz, P.C. (2000) Implementation of a broiler welfare audit to industry. South Australian Pig and Poultry Fair, pp. 42.

Barnett, J.L., Almond, A. and Glatz, P.C. (2000) A welfare audit for the chicken meat industry in South Eastern Australia. South Australian Pig and Poultry Fair, pp. 41.

Barnett, J..L., Glatz, P.C. and Almond, A. (2000) A welfare audit for the broiler industry. Proceedings Australian Poultry Science Symposium, 12: 213.

Barnett, J..L., Glatz, P.C. and Almond, A. (2000) A welfare audit for the broiler industry: A project in progress. Poultry Information Exchange, (Poultry Information Exchange Organising Committee, Caboolture, Queensland), pp. 195-196.

Barnett, J.L., Glatz, P.C. and Almond, A. (2001) Welfare standards comprehensive welfare audit for the Australian broiler industry's quality assurance program. Proceedings Australian Poultry Veterinary Association, pp. 5-6.


Barnett, J.L., Glatz, P.C. and Almond, A. (2001) A welfare audit for the broiler industry: pick-up, transport and processing. Final Report to the Rural Industries Research and Development Corporation, project DAV-162A.

Barnett, J.L., Glatz, P.C., Almond, A., Hemsworth, P.H., Cransberg, P.H., Parkinson, G.B. and Jongman, E.C. (2001) A Welfare Audit for the Chicken Meat Industry. (Rural Industries Research and Development Corporation, Canberra, Australia).

Barnett, J.L., Glatz, P.C., Almond, A., Hemsworth, P.H., Cransberg, P.H., Parkinson, G.B. and Jongman, E.C. (2001) A Welfare Audit for the Chicken Meat Industry. (Rural Industries Research and Development Corporation, Canberra, Australia) (CD-rom).

Barnett, J.L., Glatz, P.C. and Almond, A. (2001) The development of a comprehensive welfare audit for the Australian chicken meat industry and its evaluation. Proceedings 6th European Symposium on Poultry Welfare, Zollikofen, Switzerland. (in press).


Environmental Management

Project Title

Conditioning and Analysis of Broiler litter to prevent fly breeding

RIRDC Project No:

DAW-94A

Researcher: Dr David Cook Organisation: Agriculture Western Australia

3 Baron Hay Court SOUTH PERTH WA 6151


Objectives

• To determine the critical requirements of the conditioning process needed to

prevent fly breeding in poultry litter. • To evaluate the agronomic performance of conditioned poultry litter (CPL)

relative to raw poultry litter and compost in crop production (pre and post-plant).

• To network key stakeholder groups to a) provide commercial input into the field research and development of the treatment process, and b) direct communication strategies to ensure industry usage of CPL.

Background

The stable fly (Stomoxys calcitrans) has become an increasingly serious economic threat to the beef cattle and horse industries around the Perth metropolitan area. Outbreaks of this pest have forced cattle and horse owners to either relocate or agist their animals away from affected areas. As few as 20 flies per animal reduces daily weight gain and disrupts marketing plans. In addition, human lifestyle has been affected in rural residential areas. Usage of poultry litter (poultry manure plus an organic sawdust base) as a fertilizer and soil conditioner in horticultural crop production is a significant contributor to both stable fly and house fly (Musca domestica L.) populations in Perth. As a consequence of the unacceptable levels of fly breeding, application of raw poultry litter to land in horticulture will be banned in Western Australia. The conditioning (ie. daily watering and turning) of raw broiler litter represents a viable, industry alternative both in terms of process requirements and cost to end-users. However, details on the conditioning process must be finalised to ensure that no fly breeding occurs when conditioned poultry litter is used as a horticultural fertiliser; and that conditioned poultry litter has agronomic benefits comparable to raw broiler litter.

Research Conditioned poultry litter (CPL) was produced over summer, winter and spring months from litter with either a jarrah sawdust, pine sawdust or chopped straw base and evaluated for its fly breeding potential and agronomic performance compared to that of raw, untreated litter.


Outcomes Broiler litter conditioned over summer for 4, 5, 6, 7 or 8 weeks was exposed to flies

in the field and resulted in an 80, 86, 92, 95 and 96% (respectively) reduction in nuisance flies respectively relative to raw poultry litter. Storage of either 4, 6 or 8 week CPL for 2-8 weeks did not result in increased levels of fly breeding. Broiler litter conditioned over winter from 4-8 weeks was exposed to flies in the field and resulted in an 80, 86, 92, 95 and 96% reduction in nuisance flies respectively relative to raw poultry litter. Storage of either 4, 6 or 8 week CPL for 2-8 weeks did not result in increased levels of fly breeding. Broiler litter conditioned over spring from 4-8 weeks was exposed to flies in the field and resulted in an 80, 86, 92, 95 and 96% reduction in nuisance flies respectively relative to raw poultry litter. Storage of either 4, 6 or 8 week CPL for 2-8 weeks did not result in increased levels of fly breeding. CPL manure has generally performed as well or better than equivalent applications of raw poultry litter in ten agronomic trials on commercial properties (involving both pre and post-plant application of CPL to vegetables (lettuce, cabbage, onions, cauliflower and potatoes), turf and strawberries. CPL can be used at half the currently accepted best practice rate (30 cu metres/ha) for raw poultry litter, providing additional nitrogen is applied (20-25kg of Agran per week). Poultry manure applications >30 cu metres/ha, did not increase yields and actually depressed yield.

Implications The agronomic advantages of CPL were demonstrated by this research. While conditioning of poultry litter had a very significant impact on the numbers of nuisance flies bred in the material (as compared to raw, untreated poultry litter), fly breeding was not completely prevented by any conditioning treatment applied.


Other

Project Title

Production of Trial Distance Learning Materials for the Poultry Industry - Stage III

RIRDC Project No:

DAN-142A

Researcher: Mr Geoff Creek Organisation: NSW Department of Agriculture

Murrumbidgee College of Agriculture PMB YANCO NSW 2703


Objectives • To produce distance education material suitable for managers of chicken meat

enterprises. • To evaluate the distance learning materials. • To arrange accreditation for the distance education course ‘Commercial Meat

Chicken Management’.

Background

The poultry industry has limited access to specialised training. Experience on the job is the predominant method of training. Lack of adequate training may leave many supervisors and managers ill-prepared to undertake the wide range of tasks which are needed in a modern enterprise. Distance learning materials allow managers and supervisors to undertake training without leaving the workplace. This project was undertaken to complete the development of all distance learning materials required to finalise the course “Commercial Meat Chicken Management”.

Research Production of these materials involved analysis of current material, confirmation of exact training needs, development of information into tables, graphs, examples and exercises, collection of suitable photographs and diagrams, and building these resources into easy to use resource packs. Information used in the material was obtained from recognised industry experts in the relevant fields and current poultry textbooks.

Outcomes All required modules of the distance learning materials have now been developed and are available through Murrumbidgee College of Agriculture. The College provides courses under the National Agriculture Training Package that has been endorsed by the Australian National Training Authority (ANTA). The training package contains a number of Poultry Production modules/competencies. Students who are assessed to have successfully completed the distance learning modules can gain credit for a number of the level three and four poultry competencies.


Implications The industry now has available to it a complete set of distance learning materials

specifically developed to improve the competencies and skills of new and existing farm and hatchery managers/supervisors. The modules have been promoted through a series of brochures that have been distributed widely around NSW in the first instance, and through other channels. Industry has already begun to adopt and adapt these materials for their own requirements. As a result, poultry supervisors now have the opportunity to undertake formal training in specific poultry topics from their place of employment. At the same time, as these materials are taken up by industry, chicken meat producers will have increased access to staff who require less time for instruction due to the availability of relevant training materials.


CHICKEN MEAT PROGRAM – RESEARCH IN PROGRESS 45


RESEARCH IN PROGRESS

Flock Health Project Title

Detection of virulent strains of Newcastle disease virus in chickens previously infected with Australian strains of the virus

RIRDC Project No:

CSA-1J

Start Date: 01/07/97 Finish Date: 30/06/01 Researcher: Dr Harvey Westbury Organisation: CSIRO Livestock Industries

Private Bag 24 GEELONG VIC 3220


Objective

• To increase the speed, efficiency and confidence of detection of virulent strains

of Newcastle disease virus (NDV) in flocks previously infected with endemic, lentogenic strains of the virus.

Current Progress

A conventional virus capture ELISA (vELISA) for Newcastle disease virus was developed using a specific polyclonal antiserum as the capture antibody and a mixture of three monoclonal antibodies as the detection system. Positive/negative levels in the test were assessed and the best target tissues in chickens infected with virulent strains of NDV determined. These were found to be bone marrow, spleen and kidney, though regular detection was also obtained from blood and lung samples. The vELISA was also able to detect virulent ND virus in the tissues of chickens immune to the disease following earlier immunisation with ND vaccine. Immune chickens were challenged with either the virulent Herts, Texas or Australian virulent ND virus strains. These virus strains could be detected in the tissues of immune chickens for up to 21 days after challenge, with most detection occurring between 4 and 10 days after challenge. The test was used on tissues of chickens naturally infected with virulent ND virus that were collected during the Australian epidemic. Results obtained from testing these samples were not as clear-cut as those obtained with tissues from experimentally infected chickens. The reasons for this difference are being examined.


Project Title

Postgraduate scholarship – Ms Louise Hilton: Therapeutic applications of cytokines in poultry

RIRDC Project No.:

CSA-10J

Start Date: 06/06/99 Finish Date: 05/06/02 Researcher: Ms Louise Hilton and Dr John Lowenthal Organisation: CSIRO Livestock Industries

Australian Animal Health Laboratory Private Bag 24 GEELONG VIC 3220


Objective

• To enhance disease resistance and vaccine efficacy in poultry by

administration of cytokine therapy.

Current Progress

Cytokines are proteins that are naturally produced by the body’s immune system immediately following an infection. Cytokines protect against disease by controlling the immune response to infection or vaccination and therefore represent excellent, naturally occurring therapeutics. Previous poultry trials conducted by this research team have shown that treatment with a cytokine, interferon gamma, led to improvements in health and resulted in improved weight gains of the order of five to ten per cent. This cytokine also helped protect birds against coccidiosis infection. It is hoped that a range of therapeutics, using various types of cytokines, can be developed to provide producers with an alternative to in-feed antibiotics. This project has focused on the cytokine, chicken interleukin-2 (ChIL-2). Biologically active recombinant ChIL-2 was produced and its effects on the chicken immune system assessed. Tools were developed to investigate the activities of ChIL-2, including monoclonal antibodies which are used in an ELISA for measuring the level of this cytokine. ChIL-2 treatment of birds resulted in proliferation of both CD4+ and CD8+ populations of T cells. This effect indicated that ChIL-2 may be able to enhance cell mediated immunity, resulting in greater protection against a variety of viral and parasitic diseases. Currently, commercial trials are underway to study the ability of ChIL-2 to enhance vaccine efficacy in protection against infectious bronchitis virus and coccidiosis.



Project Title

Molecular epidemiology of Newcastle disease virus in Australia

RIRDC Project No:

CSA-11J

Start Date: 01/06/99 Finish Date: 30/06/02 Researcher: Dr Allan Gould Organisation: CSIRO Livestock Industries



Objectives

• To develop a better understanding of the epidemiology of Newcastle disease

virus (NDV) within Australia. • To develop a better understanding of the mutation rates as well as disease

potential of Australian NDV isolates at the molecular level.

Current Progress

Investigation of isolates associated with the outbreaks of Newcastle disease in NSW between 1998 and 2000 have shown that the progenitor virus for the outbreak arose, not from and exotic incursion of virulent virus, but from mutation of an avirulent, Australian precursor virus. Gene sequence and phylogenetic analysis of Australian NDVs isolated from 1932 to 2001 has identified one locus as a predictor of viral lineage and the likely ancestor virus for the progenitor virus has been identified. Viruses involved in the summer respiratory disease syndrome have been identified as belonging to two separate clades and virulent virus has been isolated from both clades. The entire genomes of eight viruses from these outbreaks have been sequenced and the genetic stability of these isolates determined. Selection pressure has been shown to be greatest for the haemagglutinin-neuraminidase, matrix and phosphoprotein genes. Analysis of the quasi-species or individual gene sequences present (at a low frequency) in field isolates has shown that virulent viruses were present in a background of avirulent progenitor virus. Variants in the fusion protein cleavage site (which determines virus virulence) have been isolated and characterised. Quasi-species analysis of virulent and avirulent plaque purified viruses have shown that two to three passages in vivo or in vitro were needed to attain the same genetic diversity as that identified in field isolates.

Studies of mechanisms for the natural selection of virulent viruses from field isolates have commenced.


Project Title

Biological control of necrotic enteritis in meat chickens

RIRDC Project No:

CSA-12A

Start Date: 01/10/99 Finish Date: 30/09/02 Researcher: Dr Robert Moore Organisation:

CSIRO Livestock Industries Private Bag 24 GEELONG VIC 3220


Objective

• To control necrotic enteritis in broiler chickens using a cost effective, user-

friendly, environmentally sustainable, biological control strategy.

Current Progress

Significant progress has been made towards demonstrating that bacteriocins (naturally occurring bactericidal proteins) have the potential to be used in chickens to kill Clostridium perfringens, the causative agent of necrotic enteritis. A series of synthetic bacteriocin genes have been constructed and expressed in Escherichia coli. It has been demonstrated that the recombinant protein from four of these bacteriocins has killing activity against C. perfringens. Four trials in chickens aimed at developing a suitable disease model for necrotic enteritis, were conducted. Although some encouraging results have been obtained further work needs to be done to develop a consistent, useable model. As a prelude to testing in chickens, one of the recombinant bacteriocin proteins was used in a mouse/listeria disease model to demonstrate greater than 99% reduction in pathogen load following treatment. Large doses of the recombinant bacteriocin protein had no deleterious effect on mice, demonstrating the safety of the product. A range of different bacteria are being considered for use as live vectors to deliver the active bacteriocins to chickens and research undertaken has shown that good levels of expression of active recombinant bacteriocin can be achieved in some of these potential vector strains.




Project Title

Postgraduate Scholarship - Jacqueline Kattenbelt: Mapping of structure-function relationships of Newcastle Disease (ND) using reverse genetics

RIRDC Project No:

CSA-13J

Start Date: 01/07/00 Finish Date: 30/06/03 Researcher: Ms Jacqueline Kattenbelt and Dr Allan Gould Organisation: CSIRO Livestock Industries



Objectives

• To develop a viable DNA construct containing Newcastle disease virus (NDV)

genome and separate clones of NDV nucleocapsid, protein (N), phosphoprotein (P) and polymerase (L).

• To establish a reverse genetics system to allow recovery of recombinant NDV. • To recover NDV mutants with substitutions in precise sites within the matrix

protein to give strictly defined molecular mutants. • To map structure-function relationships of viral proteins within infected cells

and to study viral morphogenesis, virulence factor and tissue trophism determinants essential for NDV replication and infection using electron microscopy.

Current Progress

Long overlapping fragments from the Peats Ridge NDV (precursor of the virulent virus which was responsible for outbreaks of Newcastle disease (ND) in Australia) have been generated and clones confirmed by polymerase chain reaction (PCR) screening and sequencing. These fragments are currently being joined at shared restriction sites to form a contiguous DNA fragment. Separate clones of nucleocapsid protein (NP), phosphoprotein (P) and RNA directed RNA polymerase (L) have been generated and cloned into expression plasmids containing an internal ribosomal entry site allowing cap independent translation. The matrix (M) gene is currently in the process of being manipulated to insert unique restriction sites to enable the gene to be cut out and a modified M re-inserted.


Project Title

Diagnostic tools for differentiation of vvIBDV and characterisation of Australian strains

RIRDC Project No:

CSA-15J

Start Date: 01/07/00 Finish Date: 30/06/03 Researcher: Dr Jagoda Ignjatovic Organisation: CSIRO Livestock Industries

Private Bag No 24 GEELONG VIC 3220


Objectives

• To introduce and compare RFLP methods developed by other research groups. • To test the specificity of Crab-88 and Crab-cos recombinant antibodies for

very virulent infectious bursal disease virus (vvIBDV) • To compare soluble, purified & phage expressed antibody for their suitability

as ELISA reagents.

Current Progress

The RFLP method was introduced and evaluated using the following overseas strains available at AAHL: classical 52/70, 1/68 and APHIS, variants E and GLS, and vvIBDV strains CS88 and Tasik. Restriction profiles generated for each virus using restriction enzymes BstN1, MboI and SspI were identical to those produced by the Jackwood and Sommer researchers who developed this method. Australian strains used in the study of Jackwood were obtained from the same source. The RFLP profile for these as well as for all other Australian strains was obtained. None of Australian strain contained an SspI site characteristic of vvIBDV strains. The specificity of the antibody Crab88 for vvIBDV strains was tested in two overseas laboratories, France and the USA. Crab88 reacted only with vvIBDV strains and did not react with any of the other classical or variant strains, including ten vaccine strains. This indicates that Crab88 is specific for all vvIBDV strains. Two other recombinant antibodies reacted with all IBDV strains, including the USA variants. Crab88 expressed as phage and soluble antibody was evaluated for use in ELISA. Higher absorbances were obtained with phage than soluble antibodies. Reagents to produce phage antibodies were cheaper and ELISA based on phage antibodies was faster and simpler.



Project Title

Evaluation of fowlpox (FPV) strains free of reticuloendotheliosis virus (REV) as vaccines for use in Australian poultry flocks

RIRDC Project No:

CSA-16A

Start Date: 01/04/00 Finish Date: 31/05/02 Researcher: Dr David Boyle Organisation: CSIRO Livestock Industries



Objective

• To undertake vaccine efficacy, safety and adventitious agent testing on

reticuloendotheliosis (REV) free fowlpox virus (FPV) strains derived from S (Standard vaccine strain) and two field strains, FPV 59vac and FPV 62vac.

Current Progress

A commercial partner for this project has been identified and an agreement has been concluded. An appropriate technical staff member has been appointed and experimental work commenced in April 2001. The first vaccination and challenge experiment has been completed. This experiment has confirmed the REV-free status of the derived strains. From the challenge component of this experiment one of the strains has been identified as suitable for further development as a commercial vaccine candidate. Master seed and an experimental vaccine stock will be produced under production conditions for this strain prior to the conduct of safety and adventitious agent testing. These tests will be conducted as a preliminary to an application for a limited field evaluation of the strain.


Project Title

Infectious proventriculitis and stunting syndrome of broiler chickens

RIRDC Project No:

DAN-171A

Start Date: 01/07/98 Finish Date: 30/09/01 Researcher: Dr Rod Reece Organisation: NSW Department of Agriculture

Regional Veterinary Laboratory Elizabeth Macarthur Agricultural Institute PMB 8 CAMDEN NSW 2570


Objective

• To understand the cause of infectious proventriculitis and stunting and to

develop relevant control strategies.

Current Progress

A few field cases of infectious proventriculitis were studied and conformed to previous descriptions. Homogenate known to induce transmissible proventriculitis was inoculated into a wide range of cell cultures, including HRT-18 (human rectal tumour) and HD-10 (chicken macrophage) cell cultures, and passaged. No cytopathic effects were noted and inoculated cell cultures were negative for immunoreactivity using convalescent chicken sera (inoculated with the same homogenate). Homogenate free of enveloped virus by chloroform treatment and free of bacteria by filtration, was inoculated into chicken embryos. The viscera of the embryos were harvested and inoculated orally into chickens but proventriculitis was not induced. Five electron-microscopic grids of transmissible proventriculitis were received from Athens, Georgia, USA, and hexagonal virus-like particles were readily observed in the nucleus and adjacent cytoplasm of proventricular alveolar epithelial cells of young chickens. At about the same time, electron-microscopic examination of proventriculi from our experimental cases at four days post-inoculation revealed similar particles. Similar particles were observed on examination of homogenate from one affected field case - this yielded reovirus on tissue culture.


Project Title

Attenuation and characterisation of chicken Eimeria for live vaccines

RIRDC Project No:

DAQ-259J

Start Date: 01/11/99 Finish Date: 31/12/02 Researcher: Dr Wayne Jorgensen Organisation: Department of Primary Industries (Qld)



Objectives

• To develop attenuated lines of E. mitis, E. brunetti and E. praecox for

incorporation in an efficacious, live vaccine, protective against all seven species of Eimeria in Australian chickens.

• To develop a trial technique to evaluate coccidiostat resistance.

Current Progress

The sensitivity of two strains of Eimeria mitis (Jorgensen and Kelly) to the coccidiostats Toltrazuril, Amprolium or Sulphaquinoxaline has been evaluated in trials. Both strains were sensitive to Toltrazuril and Amprolium. Each has since undergone selection for precocious development by passage (Jorgensen strain: 9 passages with a 20 hour drop in prepatent period; Kelly strain: 13 passages with a 14 hour drop in prepatent period). The Jorgensen strain was chosen for further characterisation studies and has been successfully tested for virulence, reproductive potential and protection against homologous challenge. A trial to assess the strain’s protection against heterologous challenge was aborted early and has now been repeated. The results of the new trial are now awaiting statistical analysis. Both strains of E. mitis have passed quality control and DNA testing for purity. The sensitivity of two strains of Eimeria brunetti (Monarto and Bowden) to the coccidiostats Toltrazuril, Amprolium or Sulphaquinoxaline has been evaluated in trials. Both strains were sensitive to Toltrazuril and Amprolium and are currently underging selection for precocious development by passage. The sensitivity of one strain of Eimeria praecox (Kelly) to the coccidiostats Toltrazuril, Amprolium or Sulphaquinoxaline has been evaluated in a trial. One potentially coccidiostat resistant isolate has been collected at Pitsworth, South East Queensland and cryopreserved for future study.


Project Title

Investigations into the development of a sustainable management strategy for the darkling beetle, Alphitobius diaperinus (Panzer) in broilers

RIRDC Project No:

DAQ-273A

Start Date: 01/07/00 Finish Date: 31/07/03 Researcher: Mr Trevor Lambkin Organisation: Department of Primary Industries (Qld)

Entomology Building, Indooroopilly Research Centre 80 Meiers Road INDOOROOPILLY QLD 4068


Objective

• To investigate sustainable management practices for the darkling beetle,

Alphitobius diaperinus (Panzer), in broiler systems which will aim at reducing pest numbers, identifying current inefficiencies in insecticide applications, sustainedly managing currently registered insecticides, developing novel control strategies and understanding better beetle population dynamics.

Current Progress

Research undertaken this last year has entailed the laboratory testing of ten east coast broiler beetle populations for resistance to cyfluthrin (Tugon®) and population dynamics and cyfluthrin efficacy studies of six south east Queensland broiler farms. Insecticide resistance test results show that mostly low levels of cyfluthrin resistance occur in beetle populations tested from Sydney, south east Queensland and the Atherton Tableland (ie less than 10% survival at the discriminating dose). Field studies of the spatial distribution of beetle populations in six clay-floored broiler sheds have found that almost all beetles that occur in the clay floors are confined to the brooder sections of the sheds. Furthermore, within the brooder sections, the majority of beetles are found under the feed pans and to a lesser extent under the drinking pans. Despite cyfluthrin showing good efficacy in laboratory tests, field studies indicate that it has no effect on beetles that harbour in the ground between clean-outs. These studies show that mortality of these beetle populations does not increase after application and that just as many live beetles occur after the application as before. In summary, results of research thus far indicate that despite many beetle populations still being susceptible to Tugon® it does little in controlling beetles that occur in clay floors. As well, if a more efficient compound is developed to treat floors it may be strategically applied to the areas only under the feed and drinking lines, which is where the majority of beetles occur.


Project Title

National NDV Survey

RIRDC Project No:

MS990-40

Start Date: 10/04/00 Finish Date: 31/07/01 Researcher: Dr Vivien Kite Organisation: Chicken Meat Program of RIRDC

PO Box 579 NORTH SYDNEY NSW 2059


Objectives

• To collect information on the type and distribution of Newcastle disease

viruses in Australian poultry flocks. • To collect information on the sero-prevalaence of NDV positive flocks across

the Australian commercial poultry indsutries and to identify risk factors for exposure to NDVs on Australian poultry farms.

• To identify possible risk factors for exposure to NDVs on Australian poultry farms.

Current Progress

The survey was designed to collect information on the sero-revalence of NDV positive flocks across the Australian commercial poultry industries, to identify possible risk factors for exposure to NDVs, and to collect information on the type and distribution of NDVs in Australian poultry flocks. The survey sampling strategy was designed to ensure comprehensive coverage of all sectors of the Australian commercial poultry industry. A total of 754 farms, across eleven regions of Australia, were sampled as part of the survey. Four types of commercial poultry enterprise were included in the survey viz. layer farms, meat chicken farms, breeder (meat and layer) farms, and dedicated pullet rearing farms. In each region, a minimum of 25% of layer farms (50% in NSW) and 30% of meat chicken farms represented in the region were included in the survey. All currently active breeder and pullet rearing farms in each region were included in the survey, except where the birds were too young. The survey was conducted in three phases. Initially, blood samples were collected and tested to establish the serological status of farms. Tracheal and cloacal swabs were then collected from seropositive farms for attempted virus isolation. Viruses isolated were genetically characterised (‘typed’), with typing based on nucleotide sequencing of the cleavage site of the gene that codes the fusion protein of NDV. A total of 259 confirmed NDV isolates were characterised, representing farms in Queensland, NSW, Victoria, Tasmania and South Australia. The majority of these isolates have come from Victorian farms. No virulent NDV isolates were detected. No precursor-like viruses (such as Peat’s Ridge or Somersby-type variants) were detected. All viruses detected have been V4-like viruses. Some minor genetic diversity was seen in these V4-like viruses,


but all are genetically distinct from the virulent virus. A full analysis of the results of the survey is currently underway.



Project Title

The development of vaccination strategies to control necrotic enteritis in poultry

RIRDC Project No:

RMI-11A

Start Date: 01/01/00 Finish Date: 31/05/02 Researcher: Prof Peter J Coloe Organisation: RMIT University

Department of Biotechnology and Environmental Biology Bundoora West Campus Plenty Road BUNDOORA VIC 3083


Objective

• To develop an effective vaccine against necrotic enteritis and to evaluate the

vaccine against a challenge model of the disease. This vaccine will be orally deliverable, cost effective to manufacture and deliverable within established farming practices.

Current Progress

Three trials have been completed with moderate success. Visible lesions were produced, consistent with necrotic enteritis, in the jejunum and ileum of birds. At post mortem, tissue from birds were given a score ranging from 0-4; 0 for no change and 4 for major tissue damage. Over the three trials the percentage of birds having a score greater than 1 from a challenge with Cl. perfringens strain 61 was: Trial 1 79% Trial 2 68% - 88% Trial 3 58% - 61% Fifty histological sections of the jejunum and ileum from challenged birds have been prepared, stained with haematoxylin and eosin and are currently being read. Some of the histological data collected from these birds suggest that challenge with Cl. perfringens may not always result in necrotic tissue manifesting as visible lesions. There may be more subtle changes of the gut that will only be detected from histological examination. The main objective for the next six months is to improve the current protocol to obtain a consistently reproducible model. To do this, the following parameters from the current protocol will be changed: diet and age of infection. A formalin-killed whole-cell Cl. perfringens vaccine will be tested on the current chicken model and assess its effectiveness in reducing mortality, severity of lesions and histological changes.


Project Title

Determination of the genomic sequence of Mycoplasma gallisepticum

RIRDC Project No:

UM-45J

Start Date: 01/06/99 Finish Date: 15/06/01 Researcher: Dr Glenn Browning Organisation: The University of Melbourne

Veterinary Preclinical Centre PARKVILLE VIC 3052


Objectives

• To determine the complete genomic sequence of Mycoplasma gallisepticum. • To facilitate identification of genes which are likely to play a role in virulence. • To lay a foundation for subsequent studies to improve the performance of

mycoplasma vaccines and to improve diagnosis of mycoplasmosis.

Current Progress

A library of clones has been constructed for sequencing. Draft coverage of the genome, with an estimated three fold redundancy, has been obtained to date. Although this data has been useful it is not in a form that can be dispersed yet, as it comprises numerous small contigs. A few gaps will remain to be closed after the completion of the random cloning phase and that is expected to take another several months to complete. The annotation will begin after that time.


Project Title

Avian Leukosis-J (ALV-J) in Australia: laboratory technologies and research needs

RIRDC Project No:

UM-49A

Start Date: 01/03/01 Finish Date: 31/07/03 Researcher: Dr Trevor Bagust Organisation: The University of Melbourne

Faculty of Veterinary Science, Pre-Clinical Centre Cnr Park Drive & Flemington Road PARKVILLE VIC 3052


Objective

• To develop the most appropriate laboratory technologies and reagents for

detection of avian leukosis subgroup J (ALV-J) and its associated disease effects for Australia's chicken meat industry.

Current Progress

Over 100 field samples have been screened for the presence of ALV-J. Such samples include vaginal swabs, whole blood and tumour tissues. Avian leukosis virus has been isolated from four tumour samples representing four different broiler breeders placed in two separate poultry operations. One additional isolate was obtained from a second Australian laboratory, where ALV-J was isolated from tumours detected in broiler breeders. The current number of ALV-J isolates being studied in our laboratory amounts to five. Identification of ALV-J has been confirmed for all five viruses using molecular-based methods. Two different sets of polymerase chain reaction (PCR) oligonucleotide primers have been used for molecular identification of ALV-J directly from tumour tissue and from infected cells (secondary chicken embryo fibroblasts) in culture. One of the PCR primer sets amplifies specifically part of the polymerase/integrase and envelope genes. The second PCR primer set amplifies specifically part of the integrase gene, the entire envelope gene, various genetic sequences located downstream of the envelope gene, and part of the 3’ long terminal repeat (3’LTR). Virus stocks have been prepared for all five ALV-J isolates, and each of the stocks is in the process of being titrated. These stocks will be used for in vivo assays and for the production of polyclonal antiserum to be used for serological assays. Sequencing of the entire envelope gene and of the 3’ untranslated region (3’UTR) is under way and the data will be used for phylogenetic comparisons of Australian isolates against published genetic sequences of foreign isolates. The sequence information will also be used to optimise molecular-based diagnostic assays.


Project Title

Control of intestinal spirochaete infections in chickens

RIRDC Project No:

UMU-23J

Start Date: 01/06/99 Finish Date: 30/08/01 Researcher: Prof David Hampson Organisation: Murdoch University

Division of Veterinary and Biomedical Sciences MURDOCH WA 6150


Objective

• To identify new and appropriate means to control infection by the intestinal

spirochaetes, Brachyspira intermedia and Brachyspira pilosicoli, recently recognised and common pathogens causing significant economic loss in Australia layer and broiler breeder flocks.

Current Progress

Studies have been conducted to determine in vitro antimicrobial drug sensitivities of spirochaete isolates from Australian chickens, and to test in vivo antimicrobial activity in experimentally infected layer and broiler breeder birds. Testing of 66 spirochaete strains revealed a similar spectrum of sensitivity to antimicrobial drugs as porcine spirochaetes. Resistance was seen against a number of drugs, with the least resistance recorded for tiamulin. In vivo, tiamulin at 25mg/kg body weight cleared experimental infection with Brachyspira intermedia in layer hens. Birds in an infected room, however, became re-infected after treatment ceased. This suggests that it may be better to use continual low level therapy to control these infections, or to use therapeutic levels of drugs combined with a thorough environmental cleaning program. Continued in-feed supplementation with zinc bacitracin at 100 ppm inhibited proliferation of B. intermedia in experimentally infected birds, whilst at 50 ppm zinc bacitracin encouraged proliferation of Brachyspira pilosicoli. It is uncertain whether these differences are due to the different spirochaete species investigated, or to the dose rates of zinc bacitracin used. Overall, these conflicting results suggest that there are complex interactions between the intestinal microflora and pathogenic intestinal spirochaete species, and that control will require careful monitoring.


Project Title

Effects of organic acids, prebiotics, and enzymes on control of necrotic enteritis and performance of broiler chickens

RIRDC Project No:

UNE-75A

Start Date: 01/10/00 Finish Date: 30/09/03 Researcher: Dr Mingan Choct Organisation: University of New England



Objectives

• To demonstrate the effect of using organic acids, prebiotics and enzymes in

broilers as alternatives to antibiotic growth promotants to maintain feed efficiency and general bird health.

• To investigate the effects of these alternative products on the prevention of necrotic enteritis.

Current Progress

The initial stages of this project concentrate on the development of a successful and repeatable disease model for necrotic enteritis (NE). Models described in the literature suggest that a dual infection of birds with Eimeria and Clostridium perfringens (CP) type A fed a diet high in wheat, fishmeal and dietary zinc will give the highest occurrence of NE in broilers. In a series of three experiments, the possibility of artificially introducing NE here at was determined. The first two experiments established that infection of three week old broiler chickens with a dosage of 7000 sporulated oocytes of Eimeria acervuline and E. brunetti will result in a subclinical coccidiosis infection with visible sings of lesions in the upper and lower intestine but only a small reduction in growth performance. A third experiment was designed to introduce NE in three- to four-week-old broiler chickens after a dual infection with coccidiosis (E. acervuline and E. brunetti 7000 oocyst each) and CP (three consecutive infections with 1ml of CP 109 CFU/ml). The result of this experiment showed that birds infected with Eimeria and CP had significantly reduced growth and feed intake and less efficient feed conversion. A future trial will have to be conducted to verify these data and identify NE on the basis of lesion scoring and bacterial enumeration.


Project Title

Enhancing mucosal immunity in chickens by novel in-ovo and postnatal vaccination techniques

RIRDC Project No:

US-72J

Start Date: 01/01/99 Finish Date: 31/12/01 Researcher: Prof Alan Husband Organisation: The University of Sydney

Dept of Veterinary Anatomy and Pathology UNIVERSITY OF SYDNEY NSW 2006


Objective

• To induce long-term immunoenhancement in chickens following early priming

of the avian immune system via in-ovo immunisation through: − development of naked DNA or recombinant constructs of antigen and/or

cytokines; − evaluation of delivery vehicles such as liposomes or biodegradable

microspheres; − assessment of immunoregulators for non-specific upregulation of the

immune system; and − evaluation of immunisation protocols for enhancing immune responses to

routine vaccinations and providing protection from disease challenges such as Salmonella Typhimurium.

Current Progress

Initial contact with and colonisation of the intestinal mucosa by pathogens is impeded by IgA antibody located at the intestinal surface. Appropriate vaccination procedures will stimulate intestinal IgA production, protecting the host from these pathogens. Such IgA antibody production can be increased through the use of immunoenhancers. Studies undertaken have investigated the immunoenhancing potential of vitamin E (VE) or the cytokine interleukin-6 (IL-6), (a communicator of the immune system, whose mammalian counterpart increases IgA production) to increase IgA antibody levels following vaccination in chickens. Dietary VE supplementation, from day old, increased antigen-specific intestinal IgA antibody titres following immunisation with either tetanus toxoid or whole killed Salmonella Typhimurium. At some dose rates, in-ovo delivery of VE with killed S. Typhimurium antigen elicited an increase (not statistically significant) in anti-S. Typhimurium IgA antibody levels post-hatch. Repeated oral delivery of IL-6 to chickens immunised with either tetanus toxoid or whole killed S. Typhimurium increased anti-antigen IgA at the intestinal surface. A live S. Typhimurium challenge model is being established to examine the ability of IL-6 induced increases in IgA antibody following immunisation, to protect chickens from a challenge of S. Typhimurium.


Bird Nutrition and Feed Supply Project Title

Characterisation of canola meal and cottonseed meal at practical inclusion levels for use in broiler and layer diets

RIRDC Project No:

DAQ-264J

Start Date: 01/07/99 Finish Date: 31/12/01 Researcher: Dr Rider A Perez-Maldonado Organisation: Department of Primary Industries (Qld)

Queensland Poultry Research and Development Centre PO Box 327 CLEVELAND QLD 4163


Objectives

• To measure the variability of glucosinolates, sinapines, condensed tannins

(CT), total phenolics (TP), sulphur and phytic acid levels in canola meal (CM) and CT, TP and free and bound gossypol levels in cottonseed meal (CSM). Pesticide residue levels in CSM; AME, amino acid and proximate composition of CM and CSM will be determined in samples from major processing sites, three times a year.

• To evaluate the ratio of iron to free gossypol in CSM to optimise production in both broilers and layers.

• To determine the upper limits of inclusion of both CM and CSM separately and in combination in broiler and layer diets.

• To make recommendations to the poultry industries on the nutritional value of both CM and CSM when included in least-cost poultry diets at levels close to their upper limit.

Current Progress

During 2000/01 several experiments were conducted to investigate the effects of CSM or CM fed at levels of 10, 20, 30 or 40% of the diet to broilers and laying hens. At 21 d chicks fed CSM showed a reduced feed intake (FI) from the 20% level. Liveweight gain (LWG) was also reduced at 20 and 30% CSM in the diet but not at the 10 and 40% levels. The feed conversion ratio (FCR) was only affected at the 20% level, where it was poorer. After 37 d LWG and FCR were not affected by any level of CSM, indicating that older birds were capable of overcoming any negative effect of CSM observed in younger birds. The results for CM indicated that after 21 d chicks fed Pinjarra CM had improved FCR at all levels, with a reduced FI but a good LWG at all but the 40% level. After 37 d a similar pattern of bird performance was observed. After 21d chicks fed Numurkah CM had a reduced FI from the 20% level but there was no effect on FCR and LWG even at the 30% level. After 37 days birds showed a similar FI and LWG response but with significantly improved FCR at all levels. After 21 d


Newcastle CM resulted in a poorer FCR from the 30% level but a good LWG and FI was observed at all levels except the 40% level. However, after 37d the birds overall FI and LWG was reduced from the 20 and 30% level, respectively, without affecting the overall FCR. After 21 d Melbourne CM resulted in a reduction in LWG and FI from the 20 and 30 % levels, respectively. However, after 37d an improved FCR was observed at all levels, even though FI and LWG were reduced from 20% level. Oil processing conditions and the presence of anti-nutritional factors may have influenced the overall performance in these meals. Although these results demonstrate that substantial amounts of CSM and CM can be used in broiler diets, more detailed studies are being undertaken to confirm the above findings. In the layer experiments, good performance was obtained when feeding CSM or different sources of CM at levels of 10, 15, or 20% of the diet. However, preliminary observations made to evaluate fresh and stored eggs derived from the above experiments indicated that an abnormal odour (fishy taint) was detected in raw eggs derived from brown layers on all CM diets. Due to the importance of this observation, further work is being undertaken using an expert sensory panel to evaluate eggs derived from hens fed CM and CSM diets.



Project Title

Estimating lysine availability by slope-ratio chick assay

RIRDC Project No:

DAQ-277A

Start Date: 01/10/00 Finish Date: 30/09/01 Researcher: Dr Rider Perez-Maldonado Organisation: Department of Primary Industries (Qld)

PO Box 327 CLEVELAND QLD 4163


Objectives

• To establish and validate a slop-ratio chick assay to determine the availability

of lysine in selected samples of canola meal and cottonseed meal. • To compare lysine availability values with ileal apparent digestibility values

determined in the same samples of canola meal and cottonseed meal.

Current Progress

Samples of canola meal (CM) from Newcastle, Melbourne, Numurkah, and Pinjarra and cottonseed meal (CSM) from Narrabri were obtained during 1999-2000 from Australian oilseed processors. Every CM and CSM samples was chemically analysed with an AME determination in broiler and layers, and a digestible amino acid determination made using broilers birds. During April-May 2001 a broiler experiment using canola meal was conducted to obtain information on the effect of formulating diets on a digestible and a total amino acid basis. The experiment was carried out in two phases, from 0-21 days (starter diets) and from 22-42 days of age (finisher diets). Dietary treatments included CM from each source which were fed at three dietary levels (20, 30 and 40%). The results of this experiment are under evaluation. A similar experiment will be conducted using cottonseed meal. It is expected that the chick assay to determine the availability of lysine in CM and CSM samples will be carried out later next year.


Project Title

Inclusion of data for additional livestock species in the Australasian Livestock Feed Ingredient (ALFI) database

RIRDC Project No:

GRD-2J

Start Date: 01/04/99 Finish Date: 30/06/01 Researcher: Dr Robert van Barneveld Organisation: Grains Research and Development Corporation

C/- Barneveld Nutrition Pty Ltd PO Box 42 LYNDOCH SA 5351


Objective

• To improve knowledge of the nutritional value of feed grains and the

efficiency of use of these grains by the egg and chicken meat industries through development of a commercial version of the Australasian Livestock Feed Ingredient (ALFI) database containing data for pigs, poultry (layers and broilers) and aquaculture species.

Current Progress

The Australasian Livestock Feed Ingredient (ALFI) database now contains more than 22,332 sample entries on the chemical composition of feed ingredients and the nutritional value of these ingredients for pigs, poultry (layers and broilers) and aquaculture species. ALFI also incorporates a vast range of information contained in recent literature and other relevant databases. The re-programmed version of ALFI has been tested by the major stakeholders and the programming of the database is now complete. A business plan has been prepared for commercialisation of the ALFI database based on distribution to end-users as a subscription service via the internet or as a CD-ROM. Final negotiations with stakeholders on commercialisation are under way.



Project Title

Premium Grains for Livestock Program (stage 2)

RIRDC Project No:

GRD-3J

Start Date: 01/07/00 Finish Date: 30/06/03 Researcher: Dr John Black Organisation: John L Black Consulting



Objective

• This project is the second stage of a major research program for improving

feed grains quality and marketing that has been negotiated in response to identified industry needs. It has five integrated component projects:

1) coordination 2) production, storage and distribution of grain samples 3) rapid and objective analytical tests for assessing feed grains quality 4) enhancing grain nutritional value through breeding and processing and 5) modelling feed grain quality.

Current Progress

Several hypotheses about the factors determining the nutritional value of cereal grains for ruminants, pigs and poultry were established in the predecessor Project GRD-1J. The relative proportion of the main chemical components of a grain is the major determinant of nutritional value for all classes of livestock. However, other grain characteristics can significantly affect energy availability and these differ between ruminants, pigs and poultry. Prediction of AME in poultry based on an assumed constant digestion of individual gross chemical components of grains show close agreement with observed values for sorghum and oats, but not for wheat or barley. The accuracy of predictions for triticale was intermediate between sorghum and barley. Further analyses showed that some of the difference between predicted and observed AME values could be explained by the viscosity of ileal digesta. However, other factors such as grain hardness and hydration capacity appear to be associated with the variation in AME between grains. In addition, starch characteristics such as granule size and distribution, amylose:amylopectin ratio and gelatinisation temperature may influence the extent of starch digestion in the small intestines of poultry and therefore energy availability. These hypotheses will be tested further in poultry over the coming year.


Project Title

Physiological limitations in energy metabolism reduce production efficiency of broilers

RIRDC Project No:

SAR-13A

Start Date: 31/10/98 Finish Date: 31/03/02 Researcher: Mr Bob Hughes Organisation: South Australian Research and Development Institute

Nutrition Laboratory PPPI, Roseworthy Campus ROSEWORTHY SA 5371


Objectives

• To develop non-invasive methods for measuring gut function in chickens. • To define the role of gut structure and function in limiting energy metabolism. • To identify the mechanism(s) by which physical and chemical properties of

feed promote sub-optimal digestion of energy. • To develop a clearer understanding of the physiological limitations of digestion

which will under-pin opportunities for development of specific strategies to reduce the cost of production of lean chicken meat.

Current Progress

Key findings to date are that gut morphology and bacterial colonisation of the gut are at least partially dependent on the sex of the chicken. Clearly, gut microflora have a highly significant impact on between-bird variation in energy metabolism in broilers. This has very important commercial implications in the nutrition and management of broilers. Sex-related differences may be important in utilisation of energy and other nutrients, in responses to anti-nutritional factors (such as non-starch polysaccharides) and various feed additives including enzymes, and in efficacy of vaccines and medication to treat gut pathogens. Up to one third of the variation in apparent metabolisable energy (AME) was associated with physical features of the small intestinal mucosa. Ileal crypt depth was the single most important feature of the small intestinal mucosa associated with variation in AME. Villus heights of the mucosa in the jejunum and ileum were significantly affected by the breed and sex of chicken, respectively. Remodeling of the villus/crypt axis, presumably in response to dietary non-starch polysaccharides in the wheat, differed in male chickens depending on breed, but there were no differences observed in female chickens. Changes in hydrogen and methane concentrations in breath during two metabolism studies were highly variable. However, observed variation between individual birds in AME was not directly associated with breath hydrogen concentration. However, elevated levels of hydrogen in breath were associated with significant reductions in growth rate and feed efficiency due to losses of energy and other nutrients through proliferation of gut bacteria.


Project Title

Improving the utilisation of dietary amino acids in meat chickens

RIRDC Project No:

US-80A

Start Date: 01/04/99 Finish Date: 31/03/02 Researcher: A/Prof Wayne Bryden Organisation: The University of Sydney

Dept of Animal Science Werombi Road CAMDEN NSW 2570


Objective

• To improve the efficiency of utilisation of amino acids in meat chickens by

formulating diets on a digestible amino acid basis; determining ideal digestible amino acid requirements; and identifying factors that influence endogenous amino acid losses.

Current Progress

Studies have been completed in which broiler diets were formulated for the growing cycle (starter, grower and finisher) using both total and digestible amino acid values. The results demonstrate clearly that using digestible amino acid values can significantly improve bird performance (growth rate, feed intake, feed conversion), breast meat yield and reduce abdominal fat pad mass. Determination of the ileal amino acid digestibility of feed ingredients used in the poultry industry is ongoing and has been extended to evaluate the effect of feed enzymes (xylanase, phytase), individually or in combination. The results suggest that the strategic dietary inclusion of exogenous enzymes will improve apparent amino acid digestibility.


Project Title

Use of dietary fatty acids to increase protein accretion in broilers

RIRDC Project No:

US-104A

Start Date: 01/02/01 Finish Date: 30/04/04 Researcher: A/Prof Wayne Bryden Organisation: The University of Sydney



Objective

• To develop a simple feed technology to reduce fat deposition and increase

muscle protein accretion in broilers by manipulating dietary fatty acid intake to alter tissue sensitivity to the metabolic hormones involved in lipid and protein metabolism.

Current Progress

Studies have commenced with fish oil (n-3 fatty acids), and sunflower oil (n-6 fatty acids) in comparison with tallow (saturated fatty acids) to determine the dietary concentration and duration of feeding required of the two polyunsaturated fat sources to change carcass composition and reduce fat deposition in broiler chickens. The studies will be extended to evaluate the effects of conjugated linoleic acid on carcass composition and performance of broilers.


Food Safety Project Title

Salmonella typing and colonisation of chickens by characterised S. Sofia

RIRDC Project No:

IMV-3A

Start Date: 01/07/00 Finish Date: 01/07/03 Researcher: Dr Michael Heuzenroeder Organisation: Institute of Medical and Veterinary Science

Infectious Diseases Laboratories PO Box 14 Rundle Mall ADELAIDE SA 5000

Phone: (08) 8222 3275 Fax: (08) 8222 3543 E-mail: [email protected]

Objectives

• To continue to provide industry with a conventional and, where appropriate,

molecular based, typing service would which will generate valuable data on the distribution of Salmonella serovars in chickens that will contribute to food safety and public health.

• To test the feasibility of AFLP typing as an epidemiological tool to replace PFGE as the preferred molecular typing method.

• To characterise by DNA sequence analysis the temperate (lysogenic) phage found in S. Typhimurium phage type 64, which is a common phage type found in chickens and humans.

• To investigate Sofia MH76 colonisation of chickens using different methods of inoculation and S. Typhimurium challenge.

Current Progress

The Infectious Diseases Laboratory has continued to receive specimens from the industry in comparable numbers to previous years. In 2000 Salmonella Sofia was the most common isolate (56.9%) from chickens. Serovars Typhimurium (20%), Kiambu (5.1%) and Virchow (4.5%) were other common Salmonella isolates. S. Sofia is still rarely isolated from humans. The PFGE (Pulsed Field Gel Electrophoresis) procedure has been shortened from six to three days. This is a great advantage as this method is used extensively in organism tracing. The AFLP (Amplified Fragment Length Polymorphism) procedure has been established in the laboratory, this method is more rapid, more discriminatory and easier to analyse than PFGE. AFLP is currently undergoing comparison with other molecular methods. The DNA sequence of two lysogenic bacteriophages ST64T and ST64B carried by S. Typhimurium PT64 has now been fully determined. These bacteriophages have


been shown to mediate phage type conversion in S. Typhimurium, which potentially may have serious epidemiological consequences. A new molecular method for typing non-typable S. Typhimurium isolates has been developed using the sequence from these phages and is yielding promising results. Colonisation studies using S. Sofia MH76 are currently being undertaken in conjunction with the Victorian Institute of Animal Science.



Project Title

Development of campylobacter bio-replacement program and establishment of campylobacter reference centre

RIRDC Project No:

UG-3A

Start Date: 01/07/00 Finish Date: 31/07/03 Researcher: Dr Victoria Korolik Organisation: Griffith University, Queensland

School of Health Sciences, Gold Coast Campus PMB 50 GOLD COAST MAIL CENTRE QLD 4217


Objectives

• To evaluate and refine the model system has been developed for colonisation

of chickens with selected Campylobacter strains. • To determine the ability of selected strains to "displace" a selection of known

campylobacter isolates that are colonising chickens and to evaluate the long-term ability of the strains to maintain a stable colonisation within a flock.

• To develop and utilise molecular methods to determine the ‘virulence potential’ of the known colonising strains.

• To further evaluate as potential bio-replacement strains for application in the chicken industry, those isolates that are shown to be negative to all identified virulence factors.

• To provide a Campylobacter reference laboratory based on molecular technology for identification of virulent and non-virulent campylobacters, which can be used for epidemiological tracing of specific strains.

Current Progress

Campylobacter spp are now the most common cause of human enteritis worldwide and it is well established that chicken meat can be a major source of human infection. Campylobacter spp also have a relatively low infectious dose and so it is not only essential that the risk of transfer of Campylobacter spp to humans via chicken meat is minimised, but it is also important to identify those strains that have greater potential to cause disease in humans. To identify the relative potential of different Campylobacter strains to cause gastroenteritis in humans, an animal model to test virulence was assessed by the intranasal infection method using the criteria of illness or death of animals as an indication of virulence. In this test, the gastrointestinal tract of 70% of inoculated mice were colonised by a known virulent C.jejuni strain, with the challenge strain also being found in the liver, spleen and lungs of some mice. However, none of the mice showed any sign of disease or discomfort. None of the other strains tested, including those isolated from patients with campylobacteriosis, could consistently colonise mice in this model. It was therefore decided to use a different strain of mice, with specific defects in mucus production in the gastrointestinal tract, which have been reported to be more sensitive to bacterial infections. This model is currently being established.


In addition, a two-day-old chick model was used to determine the colonisation patterns of the C. jejuni strains. There were five different colonisation types observed, viz. (i) immediate colonisation and prolonged excretion of viable C. jejuni bacteria; (ii) delayed colonisation and prolonged excretion of viable C. jejuni after several days; (iii) immediate colonisation with slowly clearing excretion of viable C. jejuni bacteria; (iv) delayed colonisation and slowly clearing excretion of viable C. jejuni bacteria; and (v) no colonisation of the intestines with C. jejuni bacteria. Colonisation type (i) and (ii) led to sustained colonisation of the intestines of the chickens. The maximum colonisation of C. jejuni strains before and following a passage in vivo was also determined. Colonisation ability of campylobacters isolated directly from chicken faeces has been shown to dramatically increase compared with laboratory strains or strains in a stationary phase. An increase in colonisation, of 1,000-10,000 fold, for each strain tested was observed after a single passage in vivo, but colonisation pattern type, such as immediate or delayed, remain unchanged.


Animal Welfare Project Title

Implementation of the RIRDC broiler welfare audit to industry

RIRDC Project No:

DAV-185A

Start Date: 30/11/00 Finish Date: 31/12/01 Researcher: Dr John Barnett Organisation: Department of Natural Resources & Environment (Vic)

Animal Welfare Centre 600 Sneydes Road WERRIBEE VIC 3030


Objectives

• To facilitate the adoption of audit procedures for welfare by 4 out of 6

companies in Victoria and 2 of 3 companies in SA. • To achieve participation rates in the audit program of 20% of growers and

transporters and 40% of breeder farms, rearing farms, hatcheries and processing plants in Victoria and SA by the end of the project.

Current Progress

This project involves implementing a recently completed welfare audit. The audit has been developed for all sectors of the chicken meat industry (hatchery, broilers, broiler rearers, broiler layers and pick-up, transport and processing) and is the first comprehensive welfare audit for any animal industry. Because it was developed collaboratively between researchers, industry, welfare groups and legislators it should provide for a period of certainty for the industry. The challenge with all such documents is to achieve industry adoption. The research team is currently working with several companies in Victoria, New South Wales and South Australia who are enthusiastic about implementing the audit, its role being one of encouragement and facilitation. A CD-rom is being prepared to promote the benefits of the audit to industry and this will shortly be sent out to all companies to use at appropriate producer meetings. An issue that has arisen with all companies is the need for a single and simple recording sheet/checklist. It is anticipated that the users of the audit will provide considerable input into redesigning company recording systems. Other issues to be addressed are a system of rewards/acknowledgement for participants and development of a method for capturing and publishing national compliance data.


Environmental Management Project Title

Sustainability improvements in the Victorian chicken meat industry (phase 1)

RIRDC Project No:

JSC-1A

Start Date: 15/04/01 Finish Date: 30/11/01 Researcher: Mr Jim Smith Organisation: James Smith Consulting

55 Sims Street SANDRINGHAM VIC 3191


Objectives

• To identify best practices in environmental, health, safety and community

liaison for chicken meat farming and provide tools to assist their implementation by growers.

• To establish an ongoing process to identify and address key community concerns and issues.

• To identify areas for research or technical trials required to resolve community concerns not covered by the implementation of current best practice operations.

Current Progress

The Victorian meat chicken industry is growing at four percent per year requiring the equivalent of approximately twenty new sheds annually. Community expectations for protection of the environment and neighbour amenity and for prescriptive controls enforced by state and local government authorities are growing. Although less than ten percent of the 220 broiler farms in Victoria have received complaints from neighbours regarding odour or other amenity loss issues, the drive for environmental improvement throughout the industry is strengthening and applications for new broiler sheds are frequently opposed. Programs used effectively in several other industries are therefore being adapted in the formation of the Chicken Care environmental improvement initiative for the Victorian broiler industry. In addition to helping meet community expectations, Chicken Care is expected to save Victorian growers over $3 million per year by reduced project delays, reduced legal and appeals costs and by the reduction of some unnecessary capital expenditures as conditions of new shed or farm permit approvals. The project will establish a management system for both broiler farms and the overall industry, which will demonstrate continuous improvement in the environmental performance of farms, identify and address broad community concerns and assist the meat chicken industry to grow without undue development constraints. Work on the project began in February 2001. A Community Advisory Panel has


been formed, met once, agreed its charter and begun identifying and advising the industry on a range of issues. A survey of grower implementation of the current Best Practice Model is underway. A database of the first thirty farm responses indicates compliance with over fifty percent of the practices and work-in-progress on a further twenty percent of the Model. The database is beginning to identify industry-wide areas of strength and areas in need of research or further support. Training workshops have been provided for a further thirty growers, bringing the total formally trained in the program to sixty percent of the industry growers. Initial inputs for an update to the Best Practice Model have been received to date from the Community Advisory Panel and from growers. Guidance notes on Best Practice methods for dead bird disposal, temporary litter stockpiling and contingency response plans have been developed and the topics for several other such notes have been identified. An initial set of eleven key performance indicators have been agreed by the Community Advisory Panel and the industry. Their collection by growers will begin in June 2001. Three of eight planned government briefings have been held so far with local Shire Councils and the Victorian EPA.


Project Title

Reduction of dust emissions from broiler and caged layer sheds

RIRDC Project No:

SAR-33J

Start Date: 01/01/01 Finish Date: 31/12/02 Researcher: Mr Thomes Banhazi Organisation: South Australian Research and Development Institute

Pig & Poultry Insititute GPO Box 397 ADELAIDE SA 5001


Objectives

• To determine the major risk factors associated with increased concentrations of

dust within, and dust emissions from, naturally and mechanically ventilated poultry houses.

• To develop strategies that will reduce both interior dust levels and dust emissions from buildings, resulting in more sustainable housing systems for egg and broiler production in semi-urban and more densely settled areas and reduced OH&S risks for staff.

Current Progress

The project is progressing well and according to the predetermined project timetable. The location of sheds and sampling points for measuring air quality in layer and broiler sheds (dust, ammonia etc) has been determined, including the time and season of sampling. Consultation is taking place to confirm these sampling points between the research team and Dr H. Takai of Denmark, an international expert in dust related issues in animal houses. Five different types of buildings will be studied, viz. naturally and mechanically ventilated layer and broiler sheds and tunnel ventilated broiler buildings. A questionnaire on housing for industry participants to complete has been developed. The first three farm visits for monitoring air quality was completed in late May 2001. Meanwhile discussion is also taking place with a SA based engineering firm to jointly design and evaluate a simple dust trap to be used at the air outlets of mechanically ventilated poultry sheds.

EGG PROGRAM – COMPLETED PROJECTS 79

EGG PROGRAM

COMPLETED PROJECTS

Implications of the Changing Economic Environment for the

Australian Egg Industry

Project Title

National industry databases

RIRDC Project No:

AEI-8A and AEI-9A

Researcher: Mr Hugh McMaster Organisation: Australian Egg Industry Association (AEIA)

PO Box 569 HURSTVILLE NSW 1481


Objectives

• To provide the industry with important statistics by the continuation of existing

databases • To analyse and interpret the outlook for the industry on the basis of more

comprehensive and timely data and therefore the opportunity to provide the industry with more relevant information on the market outlook

• To collect and collate information of national relevance to the Australian egg industry

• To disseminate information collated to egg producers, marketing organisations, egg products manufacturers and government personnel

Background

The Australian egg industry has few available resources for the development of nationally based statistics. The manner in which egg production is planned and the inelastic demand for eggs means the industry is extremely vulnerable to volatile swings in profitability. The development of statistics will assist in providing a better understanding of the market environment. This will assist in the development of appropriate strategic responses through revised production planning aimed to increase industry profitability.

Outcomes Improved industry statistics due to the continuation of existing databases and the production of an annual statistical publication. Also the opportunity to analyse and interpret the outlook for the industry on the basis of more comprehensive data and, therefore, the opportunity to provide the industry with more relevant information on the market outlook.


Benefits It is expected that the entire Australian industry will benefit from these databases. This is because information to be collected is considered to be relevant to the industry and is capable of regular dissemination. Possible indirect benefits may arise on the basis that a more profitable and less volatile industry may create a more certain investment climate and, thereby, facilitate adoption of more environmentally sustainable production methods.

Communication The chick placements data was communicated to the industry on a monthly basis through the RIRDC – Egg Program newsletter “Focus on Research”. Analysis of the economic outlook was presented verbally to the industry at a series of meetings throughout the country. The annual statistical publication and the final report for the project will be made available as RIRDC publications.



Project Title

Review of information sources used by Australian rural industries and egg industries overseas

RIRDC Project No:

IMS-3A

Researcher: Mr John O’Connor Organisation: Industry Management Services Pty Ltd

Level 2 470 Collins Street MELBOURNE VIC 3000


Objectives

• To prepare a review of information sources used by rural industries in Australia

and by egg industries overseas. • To prepare a presentation and attend an industry databases workshop to be

held in Sydney on 7 May 2001.

Background

As with other agricultural industries, the egg industry needs certain statistical collections for the purposes of business planning and policy analysis. There is a particular need for statistics that assist with production forecasting because egg price fluctuations are mainly caused by fluctuations in supply, not fluctuations in demand. The purpose of this project is to provide some of the information that will assist the industry to make decisions about the statistical collections to be made in the future, and the methods to be used to collect and publish those collections.

Research The main part of the report is a comprehensive listing of statistics collected for the Australian pig, dairy and wool industries, and for the egg industries of New Zealand, the UK, the USA and the Netherlands.

Outcomes The above listed examples confirm that the Australian egg industry compares unfavourably with other industries in the extent of its statistical collections and the availability of economic information such as farm physical and financial data. In the UK, the USA and the Netherlands, the egg industries are supplied with comprehensive statistical services by government agencies. In the UK and the USA, these are supplied entirely at the governments’ expense. In the Netherlands the cost is split between the government and the industry. In addition to the statistical collections, there is a substantial amount of information about farm costs, returns and performance benchmarks in these three countries. In New Zealand, the situation is much like that in Australia, with minimal contribution from the government and a very limited range of collections. The other Australian industries reported also have more extensive collections than the egg industry. To some extent this is because they are treated more generously by government agencies, but the main reason is that these industries organise and fund collections from their own resources. In some circumstances, valuable statistical series can be collected at moderate cost by using a properly constructed sample survey rather than a full survey of firms in


the relevant sector.

Implications This report provides a comprehensive illustration of the options available as demonstrated by other industries. The next, and more difficult stage, is for the industry to make judgements about the purposes for various collections and whether the cost can be justified in terms of the benefits they might confer.


Flock Health and Disease Management

Project Title

Layer industry disease and management survey

RIRDC Project No:

BIR-1A

Researchers: Dr Peter Groves Dr George Arzey Organisation: Birling Avian Laboratories NSW Agriculture

642 Great Western Highway EMAI PENDLE HILL NSW 2145 MENANGLE NSW 2568

Phone: (02) 9842 1105 (02) 4640 6402 Fax: (02) 9688 4015 - Email: [email protected] [email protected]

Objectives

• To provide prevalence information on diseases present in the layer industry. • To look for epidemiologic associations between disease and

management/environmental factors. • To look for correlations between disease prevalence, stress, performance and

management factors. • To provide industry with a better appreciation of the value of performance

recording, disease monitoring and interactions of management practices and flock health and welfare.

Background

This survey was conducted over a two year period (July 1995 through June 1997) across the layer industry in NSW and Victoria. Particular disease agents chosen for attention were Mycoplasma gallisepticum (MG), M. synoviae (MS) , Infectious Bronchitis virus (IBV) and Avian Encephalomyelitis virus (AEV). Attempts at assessing stress levels existent in flocks were also included within the survey’s data collection (differential white blood cell counts and the heterophil:lymphocyte ratio). Information was also collected on nutritional specifications, lighting programs, coccidiosis control, feed and water space allowances, vaccination histories, cage stocking densities, bird breed, beak trimming severity, egg production and mortality.

Research A random selection of 24 farms across NSW and Victoria was made, stratified across geographic locations. The farms selected were visited 3 times over a twelve month period and up to 5 age groups were examined on each visit on each farm. Each visit consisted of a clinical assessment of flock health and individual bird weights were measured on 30 to 60 birds in each age group. 12 blood samples were collected each time and tested for antibodies to MG, MS, IBV and AEV and differential white blood cell (WBC) count. If clinical disease signs were present, samples for other pathogens as deemed appropriate were also collected. A pooled faecal sample was collected and examined for presence of helminth eggs or coccidia. Cage dimensions were measured, feed and water space recorded and light intensity at the feeder trough level measured. Visual estimates of the severity of beak trim, the level of feather cover and findings from palpation of the ribs were recorded on each bird weighed. A visual estimate of shell quality (colour and presence of abnormalities) was made at each visit. A questionnaire covering feed specifications,


vaccination history, egg production and mortality history was completed over the 3 visits to each farm.

Outcomes Descriptive statistics on many factors were obtained. The period of study was notably affected by a number of epidemics, Marek’s Disease (MD) and Egg Drop Syndrome ’76 (EDS) in particular. Performance differences between the existing local breeds and the more recent imported brown egg breeds were observed. Differences in body weights of these breeds highlighted the importance of attention to welfare codes based solely on floorspace per bird rather than accounting for differences in weight per unit of floorspace. Differences in serological responses of the breeds were also shown. Some differences in performance between different production systems were also seen. Numbers of birds per cage had little influence on feather cover, shell quality or heterophil:lymphocyte (H:L) ratios. Floor production systems (free range or barn lay) elicited higher H:L ratios than did cage systems, possibly indicating higher stress levels in the former management systems. Rib scores (a measure of osteoporosis) rose with age indicating skeletal calcium depletion with length of laying period. Serological results indicated that MG entered flocks consistently in the early lay period, at a time when deleterious effects of infection could be considered to be the most serious. Many farms were encouraged to adopt MG vaccination programs as a result. IBV titres rose consistently during the laying period to a peak around 30-36 weeks of age indicating the presence of disease challenge in lay. The introduction of continual IBV vaccination was an obvious recommendation from this finding. The pattern developed for AEV serology raised concern as many farms do not vaccinate for this disease believing natural exposure in rearing provides protection. Many flocks were found to seroconvert to AEV during lay. Recommendations for proper protection by appropriate AEV vaccination was an obvious outcome. Associations between protein and calcium levels in some rearing rations with later production parameters (peak production and persistence of lay) were found. Cage space was found to be associated with flock uniformity and osteoporosis (as measured by rib palpation), surprisingly more space per bird giving poorer results. Conversely, higher weight to space ratio was associated with poorer feather cover. More severe beak trimming was associated with a later onset of lay and on an individual cage basis, more severely trimmed birds were lighter than their cage mates. Stress levels (H:L ratio) and poor feather cover were shown to be associated leading to an hypothesis that poor feather cover may be a strong contributor to stress in layer chickens.

Implications The survey provided backing for the need for importation of new vaccines (Marek’s Disease and EDS’76) to cope with the widespread outbreaks that occurred during the study period. The importance of considering bird weight in relation to cage density regulations for bird welfare was highlighted. The variances in serology and haematology may reflect a higher or more consistent exposure to pathogens with birds kept on the floor. The need for proper attention to vaccination was highlighted, especially in relation to



the patterns seen with MG, AEV and IBV in layer flocks. Cage density appears to have considerable effects, with more room per bird being deleterious in regard to egg production, uniformity of body weights and osteoporosis. Consistency and severity of beak trimming appears to have enormous effects on bird welfare and commercial performance. Due attention to the quality of this procedure would give enormous benefits to both concerns and should be given the highest priority of any bird management practice. The industry needs to recognise that poor feather cover is a major welfare issue, as well as being associated with poorer performance. This area needs much more concentrated research to understand the factors involved.

Publications Publication of results has been concentrated on feed-back to the industry: Layer disease and management survey. Scientific Meeting of the Australian

Veterinary Assoc., Poultry Information Exchange, Surfer’s Paradise, April, 1996. Survey on poultry health and management in the layer industry. Queensland Poultry

Science Symposium, Vol 5. University of Qld. Gatton, July, 1996. Factors affecting osteoporosis as measured by rib score in the layer industry survey.

Victorian Layer Production Seminar, VIAS, Attwood, Victoria, September 1997. Layer disease and management survey 1995-97: lifting our game. Poultry

Information Exchange, Surfer’s Paradise, April, 1998. Layer disease and performance survey - infectious bronchitis. Victorian Layer

Production Seminar, VIAS, Attwood, Vic, September, 1999. Layer industry survey - stress management in layers. Victorian Layer Production

Seminar, VIAS, Attwood, Vic, September, 2000.


Project Title

Revision of the AUSVETPLAN Disease Strategy document on very virulent Infectious Bursal Disease

RIRDC Project No:

BTT-2A

Researcher: Dr Clive Jackson Organisation: Biological Technology Transfer

2 Victory Avenue CAMDEN NSW 2570


Objective

• To revise the AUSVETPLAN Disease Strategy document on very virulent

infectious bursal disease.

Background

In 1998, Dr Jackson developed an AUSVETPLAN Disease Strategy Document as part of a consultancy to the DPIE. That Strategy Document was developed because of the serious economic impact that very virulent Infectious Bursal Disease (vvIBD) would have on the poultry industry should it gain entry into Australia. The annual loss was estimated to be in the order of $50 million. A revised Strategy Document was developed using the Newcastle Disease (ND) Strategy Document as a model. However, experiences gained through the implementation of that document to the 1998-2000 outbreak of virulent ND in NSW resulted in further revision of the Disease Strategy document for vvIBD.

Research The revision was undertaken with the assistance of a corresponding committee of Australian scientific experts on IBD. The Strategy Document was rewritten by the Principal Investigator to incorporate decisions from a Government/Industry meeting held on 19 August 1999. The revised Strategy Document also considered the role player by government and industry during the 1998-2000 eradication of virulent Newcastle disease in NSW. It also considered the new cost-sharing agreement for exotic disease control and eradication.

Outcomes A revised Disease Strategy document has been developed for review by RIRDC Egg and Chicken Meat Programs. The revised Strategy Document should provide the industry and government with a contemporary document on which to base an eradication program. The Strategy Document takes into account the current funding arrangements for exotic disease control in Australia. It attempts to define the resources required by industry and government. It also provides industry with guidance in undertaking a risk analysis of the benefits to be derived from pursuing eradication with limited resources and funding.

Implications The existence of a Disease Strategy document focuses the attention of the industry on the need to remain vigilant against the entry of vvIBDV. It emphasises the need to continue to develop rapid diagnostic tests to detect the viruses as early as possible. It also demands the availability of funds and resources to undertake the eradication program.



Publications Jackson, CAW (2001) AUSVETPLAN Disease Strategy for Very Virulent Infectious Bursal Disease virus (vvIBDV) Eradication. Proc. AVPA Scientific Meeting 6-7.2.01 University of Sydney, p15.

Jackson, CAW (2001) A Disease Strategy for the Prevention and Eradication of Very Virulent Infectious Bursal Disease Virus (vvIBDV). Proc. XIIth International Congress of the WVPA, Cairo, 17-21.9.01 (Abstract) (Submitted for publication).


Project Title

Postgraduate Scholarship – Matthew Rudd: Identification of virulence determinants of infectious bursal disease virus (IBDV)

RIRDC Project No:

CSA-4J

Researcher: Mr Matthew Rudd and Dr Jagoda Ignjatovic Organisation: CSIRO Livestock Industries



Objectives

• To characterise and sequence a very virulent (vv) exotic strain of infectious

bursal disease virus (IBDV) and identify genomic region(s) which may influence pathogenesis.

• To compare and contrast the pathogenicity of attenuated and very virulent IBDV (vvIBDV) strains in vivo, and identify criteria which might differentiate between the two strains.

• To establish a reverse genetics system for the recovery of infectious IBDV from cell culture, embryonating eggs and/or directly from SPF chickens.

• To swap genomic material between attenuated and vvIBDV strains and assess the impact of such manipulations on pathogenicity in vivo.

Background

Since first reported in 1989, vvIBDV has spread rapidly throughout Europe, Asia, and many other countries. Australia is currently free of such strains. This project seeks to identify potential virulence markers of IBDV for the development of rapid and highly accurate diagnostic tests which could be used to detect any incursion of exotic vvIBDV strains, should this ever occur.

Research An Indonesian vvIBDV strain was completely sequenced and the deduced amino acid sequences were aligned with the corresponding sequences of published IBDV strains to identify amino acids which are conserved solely in very virulent strains. An endemic attenuated IBDV strain (002-73) and an Indonesian vvIBDV strain (Tasik94) were both extensively characterised in vivo, and the ability of several biological assays to differentiate between the two strains were assessed. Genomic material, corresponding to a portion of the VP2 protein, was swapped between the attenuated 002-73 and very virulent Indonesian Tasik94 strains to generate recombinant IBDV. A reverse genetics system will be used to recover infectious recombinant virus. Pathogenicity testing of the recombinant virus will be used to assess the significance of putative virulence determinants identified and to provide valuable information which will assist in the development of a diagnostic assay.

Implications Further work needs to be conducted to recover and assess recombinant virus(es). The successful identification of virulence determinants is a prerequisite for


developing diagnostic tests needed to monitor the field situation of IBDV in Australia.

Publications Rudd, M.F., Heine, H.G., Parede, L., Sapats, S.I., Ignjatovic, J. (2001) Characterisation of an Indonesian strain of very virulent infectious bursal disease virus (vvIBDV). Proc. Int. Symp. Infectious Bursal Disease Virus and Chicken Anemia. In press.

Currently in preparation: Rudd, M.F., Heine, H.G., Middleton, D., Lowther, S., Ignjatovic, J. Differentiation

between endemic and exotic strains of infectious bursal disease virus (IBDV). Poster in preparation for presentation at the 3rd Veterinary Virology Conference, 26-28th September 2001.

Rudd, M.F., Heine, H.G., Sapats, S.I., Ignjatovic, J. Identification of putative virulence determinants of infectious bursal disease virus (IBDV). Manuscript in preparation for submission to Virus Research.


Project Title

Therapeutic applications of chicken interferon gamma in poultry

RIRDC Project No:

CSA-7J

Researcher: Dr John Lowenthal Organisation: CSIRO Livestock Industries



Objective

• To enhance disease resistance and vaccine efficacy in poultry by administering

therapeutic doses of chicken interferon-gamma to commercial broilers and layers.

Background

Safe, natural alternatives to antibiotics for the maintenance of optimal growth rates and flock health in chickens are being sought by the poultry industry worldwide. Cytokines are natural proteins that are produced by the body’s immune system during infection. Their ability to protect against disease make them excellent candidates as naturally occurring therapeutics.

Research Previous studies on chicken interferon gamma (ChIFN-γ) identified it as having therapeutic potential. In this project, the ability of ChIFN-γ to act as a growth promoter and vaccine adjuvant was assessed in trials using broiler chickens under commercial conditions.

Outcomes An Escherichia coli expression system was developed for the large scale production of recombinant ChIFN-γ protein. The recombinant ChIFN-γ was found to be biologically active. Monoclonal antibodies were produced and an ELISA was developed for the detection of ChIFN-γ. Treatment of broilers with ChIFN-γ resulted in enhanced weight gain over a period of up to eight weeks compared to control birds. ChIFN-γ treatment also reduced weight loss suffered by birds following infection with Eimeria acervulina.

Implications Rapid transfer of this technology to the Australian poultry industry is anticipated. These results show the feasibility of using cytokines as natural therapeutics and adjuvants. Additional cytokines have recently been identified, including interleukins -2, -6, -15 and -18. These are now available for a similar type of assessment. The particular activities of these new cytokines make them attractive new treatments for diseases such as coccidiosis, Marek’s disease and infectious bronchitis.

Publications Lowenthal, J.W., O'Neil, T.E., Strom, A.D.G., and Andrew, M.E. (1999) Cytokine therapy: a natural alternative for disease control. Vet. Immunol. Immunopathol. 72, 183-188.


Lowenthal, J.W., Lambrecht, B, van den Berg, T.P., Andrew, M.E., Strom, A.D.G., and Bean, A.G.D. (2000) Avian cytokines – the natural approach to therapeutics. Dev. Comp. Immunol. 24, 355-365.

Lowenthal, J.W. (2001) Therapeutic applications of cytokines - what can the chicken teach us? Avian Dis. (in press).

Lowenthal, J.W., Richards, G.G., Bean A.D.G., O’Neil T.E., Hilton L.S., Tyac S., Pooley, C., and Johnson M.A. (2001) New vaccination strategies for control of coccidiosis. Avian Dis. (in press).

Hilton, L.S., Bean, A,G.D., and Lowenthal, J.W. (2001) Recent advances in avian cytokines. Vet. Immunol. Immunopatol. (Invited Review – in press).

Lowenthal, J.W. et al. Chicken interferons: natural therapeutics in poultry. Avian Immunology Research Group Meeting, Turku, Finland 1998.

Lowenthal, J.W. Cytokine therapy – a natural alternative for disease control. 4th Asia Pacific Health Conference, Melbourne, 1998.

Lowenthal, J.W. Cytokine therapy – a natural alternative for disease control. 5th International Veterinary Immunology Symposium, India, 1998.

Lowenthal, J.W. Practical applications of chicken cytokines. 11th Australian Poultry and Feed Convention, Gold Coast. 1999.

Lowenthal, J.W. et al. New strategies for enhancing immunity to Eimeria: interferon gamma is the natural approach to disease control. Vaccines against Coccidioses conference, Dublin, 2000.

Lowenthal, J.W. Therapeutic applications of cytokines - what can the chicken teach us? Avian Immunology Research Meeting, Cornell University, USA, 2000.

Lowenthal, J.W. et al. New vaccination strategies for control of coccidiosis. Avian Immunology Research Meeting, Cornell University, USA, 2000.

Lowenthal, J.W. et al., Cytokines are the next generation of therapeutics. AVAP Meeting, Attwood, Vic. Oct 2000.


Project Title

NDV vaccination strategies aiming to induce high HI titres in elite breeding and layer flocks

RIRDC Project No:

DAN-159J

Researcher: Dr George Arzey Organisation: NSW Department of Agriculture

Elizabeth Macarthur Agricultural Institute (EMAI) PMB 8 CAMDEN NSW 2570


Objective

• To identify strategies capable of producing the highest and most persistent mean

HI titres in vaccinated flocks.

Background

The recent availability of inactivated ND vaccines in Australia has broadened the scope for effective long-term protection of birds against Newcastle disease. However, no sound data was previously available on the HI titres elicited when V4 was used as the priming vaccine. The present study was undertaken in order to assess a range of vaccination strategies in caged layers with the use of V4 as the priming agent

Research Ten different vaccination strategies were evaluated by monitoring the NDV HI response over a period of 28 weeks in serologically negative 18 weeks old pullets housed in commercial cages.

Outcomes Any of the combinations of live V4 plus inactivated vaccine trialled should protect a flock against clinical Newcastle disease. For protection against a drop in egg production, the strategies in which V4 was followed by inactivated vaccine four weeks later can be considered protective up to 24 weeks post initial vaccination. Protection against infection for up to three months (as determined by reduced shedding of a virulent virus) can be expected with V4 delivered orally followed with inactivated vaccine four weeks later. The studies also confirmed the poor ability of V4 to spread in flocks.

Implications The study demonstrated that V4 used as a priming vaccine with a selective combination of an inactivated vaccine can produce high and persistent NDV HI titres that are comparable to those elicited by other live overseas vaccines in combination with inactivated vaccines.



Project Title

Investigations into the management of the darkling beetle, Alphitobius diaperinus (Panzer)

RIRDC Project No:

DAQ-244J

Researchers: Mr Trevor Lambkin and Mr MC Cameron Organisation: Department of Primary Industries (Qld)

Farming Systems Institute 80 Meiers Rd INDOOROOPILLY QLD 4068

Phone: (07) 3896 9434 Fax: (07) 3896 9446 Email: [email protected], [email protected]

Objectives

• To review the relevant literature pertaining to A. diaperinus research and

thereby develop an improved understanding of the ecology of the pest. • To develop an insecticide resistance testing method for A. diaperinus and

subsequently survey and test broiler shed and egg barn pest populations in south east Queensland for insecticide resistance.

Background

The darkling beetle, Alphitobius diaperinus (Panzer) is a common cosmopolitan insect pest of poultry houses, in particular broiler sheds and egg barns, and is capable of transmitting a large number of poultry diseases and parasites. In recent concern has been expressed about increasing beetle numbers in broiler sheds and the pest’s potential to breach farm bio-security. In spite of the occurrence of the pest on almost every Australian poultry farm, no previous Australian research has been undertaken to determine the pest’s behaviour and its insecticide resistance status. Research commenced in 1998 to address these gaps in our understanding of the pest; in particular, to develop a better understanding of the ecology of the pest and to determine if resistance to fenitrothion (Folithion®) and cyfluthrin (Tugon®) has occurred in pest populations.

Research A review of scientific literature on darkling beetle research was undertaken to provide improved knowledge of the pest’s ecology, published research results and possible control strategies. Work was undertaken towards the development of an effective and efficient laboratory culture method for A. diaperinus, in order to provide a constant and adequate supply of beetles for laboratory research and testing with insecticides. A laboratory insecticide-resistance testing method was developed to provide the tools necessary to identify and characterise any insecticide resistance in A. diaperinus populations. A survey was undertaken of local broiler shed and egg barn beetle populations and these populations were subsequently tested with fenitrothion (and some with cyfluthrin). Each population tested was compared to a susceptible reference population and from this comparison a level of resistance was determined.

Outcomes The scientific literature review and project field studies have indicated that the pest’s ability to avoid contact with insecticides contributes to A. diaperinus control failures. This behaviour, together with predominantly clay-floored sheds in most broiler sheds contributes to problems with control after clean-outs, as many individuals stay concealed in the floor and do not receive a lethal dose of insecticide. The


development of a laboratory culture method has provided adequate numbers of some test insects. Problems with the culture method arose during the latter part of the project due to mite infestations, and these have hindered the availability of insects for testing. The development of fenitrothion and cyfluthrin resistance testing methods has been successful. Test results have shown that insects from south east Queensland broiler systems have strong fenitrothion resistance and some cyfluthrin resistance, and preliminary results indicate that populations of A. diaperinus from some production areas, for example Armidale and the Atherton Tablelands, have quite weak fenitrothion resistance. Insecticide resistance levels in insects from other intensive livestock systems, including egg barns are generally weaker, and all levels of resistance are directly related to duration and frequency of insecticide use.

Implications Insecticide resistance is not the major factor that determines beetle population sizes in broiler sheds. There is no relationship between anecdotal estimates of broiler beetle numbers and fenitrothion and cyfluthrin resistance levels, ie- population sizes of insects in different broiler sheds, with similar levels of insecticide resistance, can be very different. Results of testing A. diaperinus from the only south east Queensland egg barn studied show that the insects are still susceptible to fenitrothion and a rotation of fenitrothion and cyfluthrin may be done on alternate clean outs. Whether these egg barn results are typical for all is not known. For broiler sheds in general it is suggested that the application of cyfluthrin may be reduced to every second clean out (part or full) or just used over the summer period. This may delay the development of stronger resistance. Preliminary results for the broiler production areas of Armidale and the Atherton Tablelands indicate that fenitrothion may be included with cyfluthrin in an insecticide rotation. In summary, as development of strong insecticide resistance in all areas is inevitable given time, a closer examination is needed of the other major factors that control population sizes in broiler sheds. When this is known, studies of currently registered insecticides, alternative control strategies and the interaction of both can be properly evaluated.

Publications Lambkin, T.A. (1998) Controlling black beetles (Alphitobius diaperinus) in chicken

sheds. Proceedings 1998 Poultry Exchange Surfers Paradise 19-21 April 1998, 33-37.

Lambkin, T.A. and Cameron, M.C. (1999) Darkling beetle control-current difficulties and future prospects. Proceedings 11th Australian Poultry & Feed Convention 10-13 October 2000, 184-192.

Lambkin, T.A. & Cameron, M.C. (2000) Darkling beetle control in Australian broilers - a new direction. Proceedings 2000 Poultry Exchange Surfers Paradise 9-11 April 2000, 97-102.


Project Title

The development of effective immunisation strategies against Marek’s disease

RIRDC Project No:

RMI-6J




Objectives

• To improve the performance of existing Marek’s disease vaccines using suitable

adjuvants. • To characterise MDV strains after adaptation to continuous cell lines. • To develop a MDV serotype 1 specific probe to use in a dot-blot hybridisation

technique.

Background

For almost 30 years, Marek’s disease (MD) has been largely controlled by the use of intensive vaccination with herpesvirus of turkeys (HVT), either alone or in combination. However, in recent years HVT vaccines have shown to be much less effective against emerging field strains of MDV of increasing virulence. Adjuvants are used to improve the immunogenicity of a vaccine without increasing the amount of infectious virus in the vaccine. An adjuvant enhancing the performance of the HVT vaccine would be a value-added benefit to a cheap and readily available existing vaccine. The adaptation of MDVs to growth in a continuous cell line could be useful for vaccine production, compared with the labour and reagent intensive CEF cultures that are currently used. Because of limitations associated with current detection techniques for MDV serotype 1 virus, a MDV1 specific probe used in a rapid identification assay which is less expensive and more specific than those currently available, would be very useful for field diagnosis and important in vaccine evaluation.

Research Commercial broiler chickens were used to assess the possible role of γ-inulin as an adjuvant to improve the efficiency of HVT vaccination against MD. Chickens were administered vaccine with or without γ-inulin using three vaccination procedures: (i) in-ovo, by Inovoject®, (ii) in ovo by hand, or (iii) subcutaneously (sc) at day old. All birds were challenged with a virulent MDV 1 challenge virus at three days of age. Effective vaccination by HVT was assessed by the development of viraemia. HVT and MDV1 were adapted to the Vero continuous cell line (Jaikumar, 2001) and were characterised by immunological and molecular techniques. A probe labelled with digoxigenin (DIG) was developed for the detection of MDV 1


by dot-blot hybridisation. The probe was labelled by the incorporation of DIG in a PCR reaction product that consisted of the amplified 132 bp repeat located within the inverted repeat long region of the MDV 1 genome. The sensitivity and specificity of virus isolation and dot-blot hybridisation were compared with PCR, which was used as the reference procedure.

Outcomes In the vaccination trial, MD was present in all treatment groups but its incidence in groups treated with γ-inulin was not significantly different from non-treated groups. Differences in the percentages of MD in groups administered γ-inulin using the Inovoject® method or sc at day 1 were also non-significant. No adverse effects due to γ-inulin were noted in any group. HVT grew more rapidly and produced more extensive CPE and higher virus yields in Vero cell lines than MDV1. When the genome of adapted serotype 1 viruses was examined, an expansion of the 132 bp DR sequence indicated that the infected cell line contained serotype 1 MDV DNA. The presence of intact DNA with a size of approximately 180 kb in both serotype 1- and HVT-infected Vero cells after isolation and characterisation indicated that whole copies of both types of DNA were present and provided further evidence for adaptation to growth of the serotype 1 virus. This is the first report of the growth of either virus in a continuous line. Highest sensitivity rates were achieved by dot-blot hybridisation using the 132 bp PCR probe, compared with virus isolation and identification by immunoperoxidase or immunofluorescence. Despite their much lower sensitivity, higher specificity was obtained by both culture detection methods than for the dot-blot hybridization

Implications This project has shown that γ-inulin did not appear to function as an adjuvant when administered with HVT vaccine. The presence of intact DNA with a size of approximately 180 kb in both serotype 1- and HVT-infected Vero cells after isolation and subsequent analysis in a pulsed-field gel indicates that whole copies of both types of DNA were present and provides further support for adaptation to growth of the serotype 1 virus, thus would be useful for new and improved vaccine production procedures. The dot-blot hybridisation technique using the DIG-labelled probe specific for MDV 1 was shown to be potentially very useful as a rapid and economical test to detect MDV.

Publications Cipriani, T. L. (2000) The development of two digoxigenin-labelled probes for the detection of Marek’s disease virus by dot-blot hybridisation. B. App.Sc.Honours Thesis.

Jaikumar, D. (1998) Propagation and molecular characterisation of Marek’s disease virus. Master of App.Sc. Thesis.

Jaikumar, D. (2001) Adaptation of Marek’s disease virus to the Vero continuous cell line. Veterinary Microbiology 70, 75-82.


Project Title

Characterisation of very virulent Australian isolates of Marek’s disease virus

RIRDC Project No:

RMI-8J




Objectives

• To collect and maintain a repository of serotype 1 strains of Marek’s disease

virus (MDV) representative of Australia and to characterise them for future reference.

• To determine optimum methods for standardising Marek’s disease (MD) vaccines.

• To prepare and maintain reference preparations of NDV for use in vaccine assays.

• To provide an independent, reliable vaccine assay facility for use by Australian vaccine manufacturers in harmony with requirements set by NATA.

• To continually maintain and supply MDV challenge preparations for use by the industry.

Background

MD is currently a significant economic problem for the Australian chicken industry. Current, locally developed vaccines appear to provide very little protection against recently isolated very virulent strains of MDV. The availability of new vaccines increases the need for a reliable assay system to evaluate their effectiveness. Previously, vaccine assays were carried out by individual manufacturers without access to standard reference preparations. A repository will allow study trends in the evolution of MDV to be studied.

Research Blood samples were collected from flocks in different parts of the country that have been experiencing MD losses. These samples were screened for the presence of serotype 1 MDV. Positive samples were stored in liquid nitrogen for future reference. In consultation with vaccine manufacturers and industry representatives, the Virology Laboratory set up a vaccine assay facility and is seeking accreditation with the National Association of Testing Authorities (NATA). During the establishment phase of the assay, a nominal cell count and virus titre, with maximum and minimum limits, were set for two reference preparations and were revised after their substantial use to monitor the stability of the assay. Studies were carried out in response to industry concerns about possible losses to vaccine potency from the widely used practice of using chilled diluent whilst administering vaccines. Another study assessed the differences between operators whilst performing vaccine assays.


Outcomes Some 300 blood samples were collected, of which 86 were positive for serotype 1

MDV. Characterisation studies did not commence within the project’s time frame. However, despite their pre-characterisation status, the samples remain as a viable repository of MDV isolates and will be available for comparative purposes in the future. A liquid overlay procedure was adopted for the vaccine assay and over 1000 assays have been conducted since the establishment of the facility in January 1998. Few manufacturers have used the facility despite its availability to all Australian manufacturers. An impending move to a new purpose-built facility and its audit, will take place before registration with NATA. The revised cell count and virus titre results indicated the stability of each reference preparation and thus validated the nominal virus titres used throughout the assay procedure. The use of diluent held at room temperature substantially decreases virus titre loss (as opposed to holding diluent at 4°C). Therefore, it is critical to maintain diluent at room temperature whilst administrating MD vaccines to maintain their potency. Differences in vaccine titre of parallel assays of two operators were non significant and, consequently, did not affect the assay result.

Publications Two presentations on the functions of the vaccine assay facility were given at meetings of the Australian Veterinary Poultry Association at Surfers’ Paradise in April 1999 and the Victorian Poultry Health and Welfare Liaison Group in September 1999.


Project Title

Development of a live attenuated vaccine for chicken anaemia virus

RIRDC Project No:

UM-37A

Researcher: Dr Glenn Browning Organisation: The University of Melbourne



Objective

• To improve control of CAV related disease in chickens by:

Performing directed mutagenesis on the genome of chicken anaemia virus (CAV) to develop attenuated mutants

Assessment of attenuated mutants of CAV as live vaccines for administration to breeder flocks, broilers and pullets

Assessment of the suitability of DNA vaccination with these attenuated mutants.

Background

The role of chicken anaemia virus (CAV) in immunosuppressive diseases has become more evident in recent years. It contributes not only to severe immunosuppression in progeny from recently exposed parents, but also to high mortality from Marek’s disease virus associated with donor progeny susceptible to CAV, both in Australia and overseas. Increasing attention is now being paid to declining immunity in donor flock dams, which result in variable to poor progeny performance. Control of disease due to CAV currently relies predominantly on controlled exposure of breeder birds to virulent virus. This strategy will not control subclinical disease that results from exposure of older birds with waning immunity, leading to an increased risk of vertical transmission. Furthermore, this method has the problem of perpetuating the presence of virulent CAV in the environment. The need for a safe immunogenic, universally available attenuated CAV vaccine is clearly evident.

Research In order to develop an attenuated vaccine suitable for administration to all birds we have produced an infectious clone of the genome of an Australian isolate of CAV and have been introducing mutations into specific regions of the gene for VP2, one of the three viral proteins. The regions targeted are predicted to play a role in the function of VP2 during viral infection.

Outcomes Thus far we have produced a series of mutant viruses and have assessed the degree of attenuation in chick embryos. All mutants were significantly attenuated in their capacity to cause lesions in the thymus, spleen and bursa of embryos. Further studies (RIRDC Project UM-55A) are now examining the extent of attenuation in day old chicks and the protection afforded by vaccination with these mutants.


The full technical report for this project is commercial-in-confidence and is not available for general publication.

Implications The feasibility of using site specific mutagenesis to produce attenuated strains of CAV has been demonstrated. Furthermore, some vaccine candidates have been produced. Further work is needed to assess the extent of attenuation achieved, and additional mutagenesis may be needed to produce a strain that is suitable for use as a vaccine. Work still needs to be done to assess the protective immunity induced by these strains, and further work is needed to assess the potential of DNA vaccination.

Publications Brown, H. K., Browning, G. F., Scott, P. C. and Crabb, B. S. (2000) Full-length infectious clone of a pathogenic Australian isolate of chicken anaemia virus. Aust. Vet. J., 78:637-640


Project Title

Postgraduate scholarship – David Witcombe: Production and characterisation of recombinant antigens of Eimeria

RIRDC Project No:

UTS-3J

Researcher: Mr David Witcombe and Nicholas Smith Organisation: University of Technology

Molecular Parasitology Unit Dept of Cell and Molecular Biology PO Box 123 BROADWAY NSW 2007

Phone: (02) 9514 4013 Fax: (02) 9514 4026 Email : [email protected]

Objectives

• To prepare recombinant versions of a putative subunit vaccine candidate from

Eimeria, the 230 kDa merozoite protein. • To explore the potential molecular relationships between this protein and

gametocyte proteins of Eimeria. • To determine the importance of glycosylation of these proteins for induction of

immunity. • To determine how the genes encoding these antigens are organised and

expressed in the genome and whether they undergo antigenic variation or diversification.

• To assess whether combination immunoprophylaxis (eg vaccination with merozoite and gametocyte antigens) is more efficacious than vaccination with antigens from one developmental stage alone.

Background

Coccidiosis, caused by the protozoan parasite, Eimeria, is a major pathogenic disease of poultry worldwide. Coccidiosis costs the poultry industry internationally over $1 billion per year. The resistance of the parasite to anticoccidials, the high cost of development of new anticoccidials and the demands by the public for chemical-free meat drive the quest for a vaccine to control coccidiosis. Young chickens are protected against Eimeria by the transfer of antibodies from their mothers into the egg yolk, which is absorbed by hatchlings. These maternal antibodies have been used to identify proteins for inclusion in a vaccine. Exploration into the molecular composition of these proteins of Eimeria will allow deduction of the minimal components required for an effective vaccine. The development of a maternally-delivered vaccine against coccidiosis will benefit the poultry industry by allowing reduced use of chemotherapy, potentially effecting a significant cost reduction and satisfying consumer demand for residue-free meat.

Research The 230 kDa merozoite antigen of Eimeria maxima has been purified and N-terminal and internal tryptic digest amino-acid sequences determined. Using these sequences, the genetic code for the protein has been determined. Expression and production of recombinant proteins are underway. The sequences of gametocyte antigens (amino acid and genetic sequences) do not


appear to be related to the sequence of the 230kDa protein. There are several potential glycosylation sites in the 230 kDa merozoite protein. Similar sequences appear to be found in several species of Eimeria. Immunogenicity and efficacy trials using purified native and recombinant versions of the 230 kDa E. maxima merozoite protein will commence shortly.

Implications The results obtained to date suggest that the 230 kDa protein of E. maxima is a stage-specific molecule (found in asexual stages only). However, the observation that this protein is present in more than one species of Eimeria suggests that it has some conserved and important function in the development of the parasite. Therefore, it is a logical target for use in a subunit vaccine against coccidiosis.

Publications Witcombe, D., Belli, S., Wallach, M. and Smith, N. (2000) Western blot analyses and purification of an immunodominant asexual stage protein from Eimeria maxima. New Zealand and Australian Societies for Parasitology, Annual Scientific and General Meeting, Wellington, September.

Belli, S., Witcombe, D., Padula, M., Wallach, M. and Smith, N. (1999) The use of SDS-PAGE and 2-D PAGE in the analysis of parasite derived antigens. Australian Electrophoresis Society Sixth Annual Conference, Melbourne, October.

Witcombe, D., Belli, S., Wallach, M. and Smith, N. (1999) Characterisation of an immunodominant merozoite antigen from Eimeria maxima. Vaccines Against Animal Coccidioses, Interlaken, Switzerland, November.


Feed Availability and Nutrition

Project Title:

Alternative protein sources for laying hens

RIRDC Project No:

DAQ-241A

Researcher: David Robinson Organisation: Dept of Primary Industries (Qld)

The Queensland Poultry Research and Development Centre PO Box 327 CLEVELAND QLD 4163


Objective

• To broaden the base of locally available vegetable protein sources that can be

profitably included in the diet of layers. A literature review of alternative protein sources was first conducted, with special reference to grain legumes and suitability for conditions in northern Australia. Suitable recently introduced cultivars of grain legumes were identified and evaluated in laying hen trials. The nutritional profiles of these legumes were established and levels of common ANFs in the legumes were determined. Implications for the industry were discussed and recommendations with regard to further development and usage were then made, including safe maximum concentrations of the legumes in layer diets.

Background

Traditional ingredients in poultry diets are forecast to be in short supply within ten years. Inevitably there will be an increase in the world-wide demand for protein feed which is expected to be met largely by legumes and canola. As total feedgrain production in Australia is forecast to decline over the medium term, there is a risk of becoming more reliant on imports. While grain legume production in Australia is increasing, most of this harvest is grown in the southern states, primarily for human consumption. It would be highly advantageous to shift some of this production northwards, both to meet the demands of the livestock industry in this region more economically and to provide more agronomic diversity in the region. Some legume varieties are well suited to subtropical regions and show promise as competitive sources of protein for livestock. Agronomists recognise that increased cultivation of grain legumes would make a valuable contribution to sustainable agriculture.

Developments A review of alternative protein sources for layers, with special consideration for grain legumes, was completed and accompanies this report. Three varieties of chickpea, two of mung bean, two of cowpea and one of lablab were selected for evaluation for laying hens, and batches of Queensland-grown grain of each variety were obtained. Nutrient analyses, metabolisable energy determinations and measurements of a range of ANFs were completed for all these materials. Three successive rounds of laying hen performance trials (each of approximately four months duration) were conducted, using IsaBrown layers. In each trial, several


cultivars were included in nutritionally balanced layer diets at a range of concentrations determined on the basis of the laboratory information and existing knowledge of layer (or broiler) responses to the species of legume being tested. All cultivars were studied in untreated form using mash diets, but the effects of steam pelleting and decortication were also investigated with selected cultivars.

Outcomes Varieties of the same legume species were nutritionally similar, except that total sulphur amino acid levels were much lower in Amethyst chickpea than in Barwon or Dooen. Trypsin inhibitor activity was higher in chickpea and lablab than in mung bean, and higher in Amethyst than in Barwon chickpea. However, bird performance appeared to be unrelated to ANF levels, which therefore did not provide a useful indicator of safe maximum concentrations of legume in the diet. Diet composition (legume type and level) did not significantly affect mortality, and reduced mean egg weight in only one case (400 g/kg lablab). In trial 1, diets containing 450 g/kg Delta or Emerald mung bean or 300 g/kg Barwon chickpea resulted in 7-9% fewer eggs, 4-5 g/d lower egg mass and 9-10% poorer feed conversion than the control diet. Bodyweight gain over the trial period was depressed by 90-150 g in four of the six chickpea treatments. Trends in the data suggested that both Amethyst and Barwon chickpea had a depressing effect on egg mass output when included in the diet at concentrations above 100 g/kg. In trial 2, Koala lablab at 400 g/kg in mash or pelleted diets resulted in markedly lower egg number, egg mass output and feed intake and poorer feed conversion than any other treatment but did not affect body weight gain. In this trial none of the chickpea (Amethyst) or mung bean (Emerald) treatments differed significantly from the mash or pellet control treatment in respect of egg number, feed intake or feed conversion. However, birds given 450 g/kg mung bean as mash produced less egg mass but gained more bodyweight than the control birds. Egg weight was 0.87 g higher when birds were fed pelleted instead of mash diets. Mung bean at 450 g/kg in the pelleted diet resulted in 14% more eggs than the control pellets. Although there were no interactions between diet form and diet composition, diets containing 200 or 300 g/kg chickpea tended to depress performance when fed as mash but not when fed as pellets. In trial 3, moderate levels of Caloona or Red Caloona cowpea (125-250 g/kg) or Dooen chickpea (175 g/kg) tended to increase egg number and egg mass output compared to the control diet or diets containing high levels (350-375 g/kg) of these legumes. Decortication of Dooen chick pea tended to adversely affect egg mass output. Body weight gain consistently declined (or body weight loss increased) with increasing dietary levels of legumes. The average yolk colour score of eggs from birds given 375 g/kg Red Caloona cowpea was substantially higher (P<0.001) than that of the control treatment. The results overall suggest that safe dietary concentrations for long term feeding of untreated grain legumes in mash diets for laying hens are (g/kg): Barwon and Amethyst chickpea 100, Dooen chickpea 175, mung bean 300, lablab <100, Caloona cowpea 150, Red Caloona 100. These levels may be increased for short term feeding or if the diet is steam pelleted.

Implications By providing information on nutrient composition, ANF levels and safe maximum dietary concentrations for a range of grain legume varieties, this research will primarily benefit stockfeed manufacturers. Mung beans were of greatest value, followed by cowpeas and chickpeas. Benefits to the egg industry will flow through mainly from the lower cost diets that will ensue from the wider variety of feed ingredients and higher usage levels of these ingredients. The performance results provided some suggestion that inclusion of certain grain legumes at low to moderate concentrations in layer diets may improve production. ANF profiles did not provide a reliable guide to maximum inclusion rates.



This research plays an important part in the promotion of locally grown products as substitutes for imported protein meals. The grain legumes studied in this project also show potential for export growth, while at the same time they will command strong interest by the livestock feed sector. Increased knowledge of the particular limitations of different species and cultivars in poultry nutrition should encourage plant breeders to apply appropriate selection pressures to further improve the varieties suited to poultry. Grain growers should be aware that Koala lablab appears to be less suitable for poultry feeding than the other grain legumes investigated in this project. The greatest impact of this project on the Australian economy is probably its influence on the reduction of imports, expansion of exports, development of local agriculture and the long-term sustainability of the grain industry. To achieve these benefits to the fullest extent it is important to do research within the poultry sector, but the direct benefits to that sector, though significant, may turn out to be comparatively small.

Publications Robinson, D. and Datugan, M.J. (1999) Untreated chick pea and mung bean in layer diets. Queensland Poultry Science Symposium 8, pp 14-1 – 14-5.

Robinson, D., Datugan, M.J., Singh, D.N. and Barram, K.M. (2000) Evaluation of untreated grain legume varieties for laying hens. Australian Poultry Science Symposium 12: 205.


Project Title

Premium Grains for Livestock Program

RIRDC Project No:

GRD-1J

Researcher: Dr John Black Organisation: John L Black Consulting



Objectives

• To identify the reasons for and magnitude of differences between grains in their

nutritional value for ruminants, pigs and poultry so that improvements in grain quality can be achieved by plant breeding and through grain processing and storage.

• To develop rapid tests, suitable for the site of grain receipt and/or use, to measure the nutritional value of grains so that they can be priced in accordance with their suitability as an animal feed.

• To develop a computer simulation model for ruminants to predict accurately the consequences of grain characteristics and of grain processing and storage on the productivity of feedlot cattle.

Background

The Program involved collaborative funding from the Grains, Pig and Dairy R&D Corporations, Meat and Livestock Australia and the Chicken Meat and Egg Programs of RIRDC. The Program is expected to improve the quality of grains available to the animal industries and to provide a more rational basis for trading feed grains based on measurement of the nutritional characteristics of grains determining their quality for different livestock enterprises.

Research Over 2000 grain samples covering the widest possible range in chemical and physical characteristics that may influence animal performance have been collected. The samples have been derived from germplasm collections, plant breeders, specially grown crops and commercial grains suspected of having extremes in nutritional values because of severe drought, frost damage or germination. All samples collected have been scanned with near infrared spectrometry (NIR). Approximately 115 analyses of chemical and physical characteristics thought to influence nutritional value have been conducted on all grains fed to animals. These involved analyses for individual carbohydrate, fatty acid and amino acid components, α- and β-amylase and anti-nutritional factors such as lectins, tannins and phytic acid. Physical properties included measurement of grain weight, hydration capacity, seed colour, seed diameter, seed size distribution, seed hardness index and profile, and the viscosity of whole grain, starch extract and acid soluble extract. Light and electron microscopy has been used to examine the physical structure of some grains that differ markedly in their nutritional value. Many grains, including all those fed to animals have been examined using in vitro systems simulating both rumen fermentation and intestinal digestion. These analyses



have been extremely useful for identifying grains that potentially have large differences in nutritional value for different classes of livestock and for screening potential processing procedures. A relatively small number of grains (approximately 100) covering the range identified in chemical and physical characteristics have been fed to sheep, cattle, pigs, broiler chickens and laying hens to determine the digestion of energy, individual grain compounds and, in some animal species, amino acids. The effects of processing and storage on the nutritional value of grain for different animal species have been evaluated using the in vitro systems. Development of rapid and accurate analytical tests for measuring the most important chemical and physical characteristics that determine nutritional value of feed grains has commenced. Preliminary analyses using NIR have been developed for predicting the digestible energy content of grains for pigs, apparent metabolisable energy (AME) for broiler chickens and whole animal dry-matter digestibility for sheep. In addition, NIR procedures have been used for predicting the major chemical components of grains and for predicting the in vitro digestion of various grain components.

Outcomes Results show that there is a wide variation in energy availability both within and between cereal grain species and between animal types. For example, the observed range in AME (MJ/kg) for broiler chickens for the grains examined in the Program is 15.4-16.1 for sorghum, 13.2-14.7 for wheat, 11.2-13.2 for barley, 11.2-14.4 for triticale, 12.6-13.4 for normal oat grain and 14.6 for naked oats. Naked oats had an AME value of 16.2 for laying hens. Sorghum has a much lower available energy content for cattle at 9.7 MJ/kg than for pigs (14.6 MJ/kg) or broiler chickens (15.9 MJ/kg). The energy value of waxy sorghum is enhanced considerable for cattle to 13.2 MJ/kg, but there was only a marginal increase of 0.1 to 0.5 MJ/kg for pigs and poultry.

Implications Several hypotheses about the factors determining the nutritional value of cereal grains for different animal types have been established and are being evaluated in the current Project GRD-3J.

Publications Balogun, R., Bird, S. H. and Rowe, J. B. (2000) Improving the nutritive value of sorghum grain by germination. Asian-Australian Journal of Animal Science. 13 (supp): 160.

Bird, S. H., Rowe, J. B., Choct, M., Stachiw, S., Tyler, P. and Thompson, R. D. (1999) In vitro fermentation of grain and enzymatic digestion of starch. Recent Advances in Animal Nutrition in Australia (Editor J. L. Corbett), 12: 53-61.

Black, J. L. (1999) Nutritional value of cereal grains for animals. Chemistry in Australia. 66: 7-9.

Black, J. L. (1999) The Premium Grains for Livestock Program. In: The Eleventh Australian Poultry & Feed Convention, Royal Pines Resort Gold Coast, Australia. pp. 226-32.

Black, J. L. (2001) Variation in nutritional value of cereal grains across livestock species. Australian Poultry Science Symposium, University of Sydney, Sydney, Australia 13: 22-9.

Choct, M., Hughes, R. J., Perez-Maldonado, R. and van Barneveld, R. J. (2001) The metabolisable energy value of sorghum and barley for broilers and layers. Australian Poultry Science Symposium, University of Sydney, Sydney, Australia 13: 39-42.

Dixon, R. M. and Stockdale, C. R. (1999) Associative effects between grains and forages: consequences for feed utilisation. Australian Journal of Agricultural Research 50: 757-73.


Evers, A. D., Blakeney, A. B. and O’Brien, L. (1999) Cereal structure and composition. Australian Journal of Agricultural Research. 50: 629-50.

Farrell. D. J. (1999) In vivo and in vitro techniques for the assessment of the energy content of feed grains for poultry: a review. Australian Journal of Agricultural Research 50: 881-8.

Flinn, P. C. (2000) Current and potential use of NIR in the fodder and grain industries: a ruminant’s perspective. In: Reading more from the spectrum, 9th Australian Near Infrared Spectroscopy Conference, Horsham, VIC, Apr 2000.

Flinn, P. C. (2000) Rapid methods for the quality assessment of grains for animal nutrition. In: Chemical aspects of grain in human and animal nutrition., Cereal Chemistry Division Symposium, 11th RACI Convention, Canberra, ACT, Feb 2000, p15.

Flinn, P. C., Heazlewood, P. G. and Dalton, S. L. (2000) Recent developments in improving the prediction of digestibility of feed grains. In: 9th International Conference on Near Infrared Spectroscopy Verona, Italy, Jun 1999 (in press).

Hogan, J. P. and Flinn, P. C. (1999) An assessment by in vivo methods of grain quality for ruminants. Australian Journal of Agricultural Research 50: 843-54.

Hughes, R. J. and Choct, M. (1999) Chemical and physical characteristics of grains related to variability in energy and amino acid availability in poultry. Australian Journal of Agricultural Research 50: 689-701.

Hughes, R. J., Choct, M. and van Barneveld, R. J. (2001) Factors influencing the energy values of Australian cereal grains fed to broilers. Australian Poultry Science Symposium, University of Sydney, Sydney, Australia 13: 30-8.

Kaiser, A. G. (1999) Increasing the utilisation of grain when fed whole to ruminants. Australian Journal of Agricultural Research. 50: 737-56.

Kitessa, S., Flinn, P. C. and Irish, G. G. (1999) Comparison of methods used to predict the in vivo digestibility of feeds in ruminants: A review. Australian Journal of Agricultural Research 50: 825-41.

Kruk, J. A. and van Barneveld, R. J. (1999) Near infra-red spectrophotometry predictions of digestible energy in cereals for pigs. In: Manipulating Pig Production VII (submitted, Editor P. D. Cranwell) Australasian Pig Science Association, Werribee, VIC.

Nagorka, B. N. (2000) A ruminant model to estimate the nutritive value of grains in cattle feedlots. International Report, CSIRO CLI.

Nagorka, B. N., Gordon, G. L. R. and Dynes, R. A. (2000) Towards a more accurate representation of fermentation in mathematical models of the rumen. Modelling Nutrient Utilization in Farm Animals (Editors McNamara, J. P., France, J. and Beever, D. E.) CAB International, London (in press).

Moughan, P. J. (1999) In vitro techniques for the assessment of the nutritive value of feed grains for pigs: a review. Australian Journal of Agricultural Research 50: 871-9.

O’Brien, L. (1999) Genotype and environment effects on feed grain quality. Australian Journal of Agricultural Research. 50: 703-19.

O’ Brien, L. (2000) Genotype and environment effects on feed grain quality. Royal Chemical Institute, 11th National Convention, Canberra, ACT, Feb 2000.

O’Brien, L. and Blakeney, A. B. (2000) Genetic variation for components of dietary fibre in wheat and barley grains. 11th Cereals and Bread Congress, Gold Coast, Sept 2000.

O’Brien, L. and Blakeney, A. B. (2000) Opportunities for genetically improving the dietary fibre content of wheat and barley. International Dietary Fibre Congress, Dublin, Ireland, May 2000.

O’Brien, L., Tredea, A. M., van Barneveld, R., Bird, S. and Rowe, J. (2000) Genotype and environment effects on measures of feed grain quality in wheat, barley and oats. 11th Cereals and Bread Congress, Gold Coast, Sept 2000.


Petterson, D. S., Harris, D. J., Rayner, C. J., Blakeney, A. B. and Choct, M. (1999) Methods for the analysis of premium livestock grains. Australian Journal of Agricultural Research 50: 775-87.

Ravindran, V. and Bryden, W. L. (1999) Amino acid availability in poultry – in vitro and in vivo measurements. Australian Journal of Agricultural Research 50: 889-908.

Rowe, J. B., Choct, M. and Pethick, D. W. (1999) Processing cereal grains for animal feeding. Australian Journal of Agricultural Research 50: 721-36.

van Barneveld, R. J. (1999). Chemical and physical characteristics of grains related to variability in energy and amino acid availability in pigs: a review. Australian Journal of Agricultural Research 50: 667-87.

van Barneveld, R. J. (1999) Controlling the nutritional quality of stockfeed ingredients. VICTAM Feed Ingredients and Grain Processing Asia ’99. Bangkok, Thailand, 3-5 November, 1999.

van Barneveld, R. J. (1999) Physical and chemical contaminants in grains used in livestock feeds. Australian Journal of Agricultural Research 50: 807-23.

van Barneveld, R. J. (1999) Strategies for the assessment of livestock feed ingredient quality. Recent Advances in Animal Nutrition in Australia 12: 63-72.

van Barneveld, R. J., Ru, Y. J., Wyatt, G. F. and Pluske, J. R. (2000) Relationship between the ileal and faecal digestible energy content of pig diets containing Australian barley cultivars. Proceedings of the Nutrition Society of Australia (in press).

van Barneveld, R. J., Ru, Y. J., Szarvas, S. R. and Wyatt, G. F. (1999) Range in digestible energy and true ileal digestible lysine content of 11 barley samples. In: Manipulating Pig Production VII (submitted, Editor P. D. Cranwell) Australasian Pig Science Association, Werribee, VIC.

van Barneveld, S. (1999) Chemical and physical characteristics of grains related to variability in energy and amino acid availability in ruminants: a review. Australian Journal of Agricultural Research 50: 651-66.

White, C.L. and Ashes, J. R. (1999) A review of methods for assessing the protein value of grain fed to ruminants. Australian Journal of Agricultural Research 50: 855-69.

Wrigley, C. M. (1999) Potential methodologies and strategies for the rapid assessment of feed-grain quality. Australian Journal of Agricultural Research 50: 789-805.

Zarrinkalam, M. R., van Barneveld, R. J., Tivey, D. R. and Choct, M. (1999 Predicting energy availability in barley for pigs and poultry using rapidly determined fibre content. In: Manipulating Pig Production VII (submitted, Editor P. D. Cranwell) Australasian Pig Science Association, Werribee, Vic.


Project Title:

Amino acid and energy requirements of imported IsaBrown laying hens

RIRDC Project No:

US-54A

Researcher: D Balnave* and D Robinson** Organisation: *University of Sydney, Department of Animal Science, Camden, NSW 2570 and

**The Queensland Poultry Research and Development Centre, Cleveland, Qld 4163 Phone: *(02) 4655 0677 **(07) 3824 3081 Fax: *(02) 4655 0693 **(07) 3824 4316 Email: *[email protected] **[email protected]

Objective

• To improve the economic value of imported commercial brown layer strains by

defining their protein, amino acid and energy requirements under Australian conditions.

Background

During the past decade egg producers in Australia have to a large extent discarded established layer strains in favour of new coloured overseas genotypes. These imported brown-egg strains produce considerably more egg mass and generally convert feed to egg mass more efficiently than local strains, so it might be expected that their nutritional requirements are more exacting than those of local strains. However, no estimates of the nutritional requirements of these “imported” strains have been made in Australia using Australian diets. Previous attempts to evaluate the performance and to determine the dietary nutrient specifications of these overseas strains in the Australian environment have been impeded by a high mortality problem related to Marek’s disease and cannibalism. The recent reduction in the incidence of Marek’s disease provided an opportunity to evaluate the nutrient requirements of these imported stock under Australian conditions.

Research Long term laying trials were conducted with IsaBrown laying hens in which variations in dietary energy, protein, lysine and methionine concentrations were examined in separate studies. Measures of production, egg quality and immunological status were determined to help identify the optimum dietary concentrations of these nutrients.

Outcomes The IsaBrown hen appeared to be rather inefficient at adjusting feed intake to meet energy requirements. Results suggested that a minimum dietary apparent metabolisable energy of 11.4 MJ/kg appeared to be required for optimal biological efficiency. This agrees with current breeder recommendations. The lysine and methionine requirements for hens in single cages were lower than for hens in multiple 5-bird cages. The requirements of the latter hens are more applicable to the commercial situation and were shown to approximate 19.5 g crude protein/day, 970 mg lysine/day and 430 mg methionine/day. At a dietary concentration of 11.25 MJ/kg these intakes would be attained with approximate dietary concentrations of 160 g crude protein/kg, 7.75 g lysine/kg and 3.33 g methionine/kg. These determined requirements for crude protein and methionine were similar to breeder recommendations but the determined lysine requirement was considerably higher (970 vs 880 mg/day).

Implications These results suggest that, for the IsaBrown laying hen, diets with a low to medium energy content supplying 970 mg lysine and 430 mg methionine daily will be

mailto:*[email protected]


satisfactory for egg production.

Publications Balnave, D., Gill, R.J., Li, Xiuhua and Bryden, W.L. (2000) Lysine dose response study with IsaBrown laying hens housed in single or multiple cages. Aust. Poult. Sci. Symp. 12: 201.

Balnave, D., Gill, R.J., Li, Xiuhua and Bryden, W.L. (2000) Responses of IsaBrown laying hens to a pre-layer diet containing additional calcium and to dietary protein and lysine concentrations during lay. Aust. J. Agric. Res.(In press).

Robinson, D., Schermer, M. and Datugan, M.J. (2000) Effects of dietary energy level and cage stocking density on performance of IsaBrown laying hens. Aust. Poult. Sci. Symp. 12: 105-108.


Husbandry and Welfare

Project Title

Development of a non-invasive test of stress in laying hens

RIRDC Project No: US-71A Researcher: A/Professor Wayne Bryden Organisation: University of Sydney



Objective

• To develop a non-invasive test of stress in laying hens for the assessment of bird

welfare. Specifically to determine if hormones released in response to specific stressors are sequestered from systemic circulation into egg albumen.

Background

Of increasing concern to the egg industry is the growing public perception that laying birds exist in a state of chronic stress for the duration of their productive life. In view of the public concern of the welfare issues associated with egg production it is important to try to identify or define what is a contented bird exposed to minimum stress.

Research In a series of studies the relationship between circulating concentrations of stress hormones (corticosterone and catecholamines), and their sequestering into egg albumen was determined under normal conditions and experimentally induced stress.

Outcomes Corticosterone levels in egg albumen increase when hens are exposed to stressors. Moreover, corticosterone is easily measured in albumen and sample processing is relatively inexpensive, both important aspects of a rapid assay method. In contrast, catecholamines are difficult to measure and no evidence was found to indicate that egg albumen levels of these hormones are increased by stressors, which influence corticosterone levels. It is concluded that catecholamine levels in albumen fail to provide a non-invasive measure of stress in hens.

Implications The project has shown that a more extensive evaluation of albumen corticosterone concentrations as a measure of stress in hens is warranted and should provide a very useful tool in assessing the effects of husbandry and housing on hen welfare.

Publications Downing, J.A. and Bryden, W.L. (1999) Stress, hen husbandry and welfare. Literature Review of Stress in Poultry, for RIRDC/Egg Program. p.58.

Downing, J.A., Fraser, D.R. and Bryden, W.L. (2001) Development of a non-invasive test for stress in laying hens. Proc.Aust.Poult.Sci.Symp. 13 (In press).

Downing, J.A. and Bryden, W.L. (2001) Egg albumen corticosterone concentration in hens exposed to high temperature. Proc.Aust.Poult.Sci.Symp. 13 (In press).



Environmentally Sustainable Management Project Title

Environmentally acceptable land use for chicken meat and egg production

RIRDC Project No:

UNE-59A

Researcher: Dr Roger Epps Organisation: University of New England

School of Human and Environmental Studies ARMIDALE NSW 2351

Phone: (02) 6773 2904, (02) 6773 3360 Fax: (02) 6773 3030 Email: [email protected]

Objective

• The main objective of this research project was to review the nature of land use

conflict on the urban fringe by focusing on the Australian poultry industries. Additional objectives included investigating how government regulates the off-site impact of poultry farms, possible implications for agricultural production and alternative conflict management strategies.

Background

The Australian poultry industries have undergone a rapid process of intensification since the 1960s. Intensification may negatively impact on neighbouring property owners, where odour, noise and other externalities are produced. Because the poultry industries have traditionally been located on the urban fringe, land use conflict has increased in recent decades as Australia’s main urban centres have expanded outwards.

Research The focus was on the urban fringe of Perth, WA and Sydney, NSW. Interviews were conducted with: poultry farmers; egg companies and chicken meat processors; farmer association leaders; local government officials and state government agencies.

Outcomes The nature of conflict was similar between Perth and Sydney, with the chicken meat industry attracting a higher level of complaint (especially odour and night time noise) and farmers facing increasing difficulty in developing new farms in outer fringe areas. Although local governments in both States had attached tougher consent conditions on shed approval, the regulatory approach to conflict varied. The WA Government and the WA broiler industry have been more proactive in dealing with conflict compared to NSW where local government initiatives have received inadequate support.

Implications The political balance in rural areas is changing such that claiming ‘existing rights’ may hold little weight when councillors are lobbied by a large number of local residents. It is critical that the industry gives greater attention to scientific research and to addressing community concerns. Local government must become more aware of conflict, because farmers may not relocate at the first sign of complaint.


Greater importance must be given to strategic planning, especially to avoid conflict simply being relocated into rural areas. State government must recognise the potential for conflict on the urban fringe, and, where necessary, provide greater support for buffer distances or explore options that may assist the relocation process. If land is to be designated for agriculture, minimum performance standards need to be established, which, if achieved, should simplify the approval process and help to avoid politically motivated decisions. If a proactive approach is adopted, then fewer resources may need to be directed to regulating complaints in the future. Alternatively, if farmers consider relocation to be difficult, in terms of obtaining approval, and financially costly, given the increasing number of consent conditions, then they may decide to continue operating on the urban fringe, while awaiting retirement – a decision that may simply prolong land use conflict.

Publications Henderson, S.R. (2001) ‘Farmer Experiences of Land use Conflict and the Role of Mediation in Conflict Resolution’, Paper presented at the joint conference of the New Zealand Geographical Society and the Institute of Australian Geographers, Univ. of Otago, Dunedin, 29 Jan - 2 Feb.

Henderson, S.R. (1999) ‘Land Use Conflict on the Urban Fringe: Definitions and Dimensions’, Paper presented at the Inst. of Aust. Geographers Conference, Univ. of Sydney, 27 Sep - 1 Oct.

Henderson, S.R. (1999) ‘Agri-food production on the urban fringe – environmental issues facing the Australian poultry Industry’, Paper to VII Agri-Food Conf, Univ. of Sydney, 15-16 July.

Henderson, S. and Epps, R. (1999) ‘Rural land use change and the ‘Ratchet’ effect’, in Geodiversity: Readings in Australian Geography at the Close of the 20th Century, Special Publ. Series No. 6, Canberra, ACT, School of Geography and Oceanography, ADFA, pp 457-466.

Henderson, S. and Epps, R. (1998) ‘Sustainability and intensive agriculture in the rural-urban continuum’, in Sustaining Rural Systems in the Context of Global Change, Proc. Conf. of the IGU CSRS and LUCC, UNE, 5 -12 July 1997, R. Epps (ed) SHES, UNE, Armidale, pp. 197-204.

Henderson, S. and Epps, R. (1997) ‘Implications of expanding intensive agricultural production: two contrasting regional studies’, Paper to, ANZRSA Conf., Univ. of Victoria, NZ, 8-12 Dec.


EGG PROGRAM – RESEARCH IN PROGRESS 115

EGG PROGRAM

RESEARCH IN PROGRESS

Implications of the Changing Economic Environment for the Australian Egg Industry

Project Title

The economic impact of changing Australian egg production systems

RIRDC Project No:

ANU-41A

Start Date: 18/05/00 Finish Date: 31/05/01 Researcher: Dr Ray Trewin Organisation: Australian National University

Australia-Japan Research Centre (AJRC) Building 13 CANBERRA ACT 0200


Objective

• The key deliverables will be the production of RIRDC and other publications,

and their wide presentation including to industry, governments and researchers. The publications will cover key issues concerning the socio-economic impact of changing Australian egg production systems such as the costs and benefits of possible future scenarios to industry, consumers, and society. The publications, which will be widely distributed, will assist Australian policy agencies and businesses to make decisions based on the full socio-economic consequences of possible changes in Australian egg production systems. The outcome of the proposed research will be more informed decisions by Australian policy agencies and businesses in relation to Australian egg production systems.

Current Progress

The project started early in mid May 2000 to enable preparation of an Issues paper for a July Hen Housing conference that was called at short notice. The paper was discussed with industry, government officials and other Australian researchers prior to the closed conference. The Issues paper was also discussed with international researchers at the International Association of Agricultural Economics Conference in Berlin in August 2000. Further discussions took place with other researchers, industry and government officials in the UK and Switzerland during a study tour following the Conference when relevant material such as the National Farmers


Union survey of producers and estimates of welfare benefits were obtained. A Management Committee was formed and met in early November 2000 to discuss a redraft of the Issues paper and to provide advice on the industry data collection stage. An extended paper was presented to the Australian Agricultural and Resource Economics Society Conference in Adelaide in January 2001. A foresighting exercise of the industry was undertaken in May before preparation of the final report with estimates of economic costs and benefits of changing Australian egg production systems for early June 2001.



Project Title

Options for enhancing industry competitiveness and R&D and marketing efficiency (EIDF)

RIRDC Project No:

AEI-11A

Start Date: 06/04/01 Finish Date: 30/11/01 Researcher: Mr Alan Newton Organisation: Australian Egg Industry Association (AEIA)



Objectives

• Enhancement of the Australian egg industry international competitiveness and

capacity to relate effectively with government. • Appropriate innovation, marketing, and policy capabilities to equip the

industry to meet the above. Specific deliverables are: • Reports to industry on analysis of options for enhancing industry

competitiveness and for delivering more efficient egg innovation, marketing, communication and policy outcomes.

• Subject to industry decisions on the above, the preparation of appropriate proposal(s) for submission to government.

• A comprehensive process of industry consultation and reporting to progress the above.

• Assistance in the full development and implementation of any formulation agreed by industry and government.

Current Progress

Project supported out-of-session. Insufficient progress to warrant a progress report at the time of preparing this publication.


New and Existing Markets Project Title

An evaluation of the higher value-added opportunities from the chicken egg

RIRDC Project No:

DAQ-275A

Start Date: 01/11/00 Finish Date: 30/11/01 Researcher: Dr Craig Davis Organisation: Department of Primary Industries (Qld)

Centre for Food Technology 19 Hercules Street HAMILTON QLD 4007


Objective

• To conduct a comprehensive desktop investigation to determine the current and

potential uses of extracts and by-products produced during egg processing.

Current Progress

Report not received.




Public Health Project Title

Rapid detection of virulent Salmonella in egg and poultry products

RIRDC Project No:

CIF-1A

Start Date: 01/07/00 Finish Date: 30/11/03 Researcher: Dr Jason Wan Organisation: CRC for International Food Manufacture and Packaging Science

Private Bag 16 WERRIBEE VIC 3030


Objectives

• To develop multiple PCR systems to target a spectrum of multiple (at least 6)

gene sequences for rapid characterisation and differentiation of Salmonella spp. of economic and public health significance in egg and poultry products and environmental samples, and correlation of the genetic information with sero-typing and phage-typing data.

• To develop simple sample preparation techniques for the isolation and concentration of Salmonella spp. from egg products for use in combination with the multiple PCR systems.

• To apply the developed detection and identification systems for differentiation of non-pathogenic S. sofia and pathogenic non-S. sofia isolates and S. Enteriditis (SE) PT4 (a major poultry pathogen in Europe & USA) and other Salmonella spp. of industry importance.

• To develop user-friendly systems such as "BioChips" (DNA micro-array systems) and colorimetric detection systems for the detection of multiple PCR products.

Current Progress

A comprehensive literature and genomic database review on virulence determinants has been completed and a reference culture collection of 54 relevant Salmonella strains has been compiled. Multiple gene sequences in the Salmonella genome have been identified as potential targets for differentiation and characterisation of Salmonella isolates. Multiple pairs of oligonucleotide primers targeting specific regions of a virulent determinant have been synthesised for PCR differentiation of Salmonella species into the major sero-groups (B, C1, C2-3 and D) and for correlation with sero-typing results. In addition, PCR systems are also being developed for amplification of common gene targets in all Salmonella species, and the PCR products are being sequenced using a DNA sequencer for analysis of S. Sofia-specific regions. The sequence information generated will fill the knowledge gaps in genetic information for local isolates of S. Sofia and will be used as additional targets for differentiation of S. Sofia from other Salmonella species.


Project Title

Egg and egg shell quality control in the Australian egg industry

RIRDC Project No:

UNE-71A

Start Date: 01/07/99 Finish Date: 31/07/02 Researcher: A/Prof Juliet Roberts Organisation: University of New England

Dept of Animal Physiology School of Rural Science and Natural Resources ARMIDALE NSW 2351


Objective

• To provide a data base and bench marks for egg and egg shell quality in the

Australian egg industry. The relationship between laboratory measurements and the commercial situation (proportion of eggs lost or downgraded before and during grading) will be established by compiling data obtained from grading floors and egg marketing organisations along with data obtained from the field and the laboratory. The results will be summarised in a "user-friendly" booklet which will provide graphs of egg and egg shell quality values for different strains of birds at different ages, under different conditions. The booklet will be marketed through RIRDC.

Current Progress

Some flocks of birds are still being analysed for the longitudinal studies, where individual flocks of birds are followed throughout their laying life. However, the emphasis of the project has now shifted to the cross-sectional studies, where flocks are sampled from eggs delivered to egg grading and washing facilities. For the longitudinal studies, eggs have been received from six locations, twenty-nine flocks representing four strains, three types of caged housing systems as well as free range, across all seasons of the year and at a total of sixteen different ages of hen. For the cross-sectional studies, eggs have been received from eleven locations, fifty-five flocks, representing four strains, across all seasons and at thirty different ages of hen. Preliminary analysis of some of the data indicates that there is a degree of variation for egg and egg shell quality measures from flocks of the same age. In order to explain this variation, data will be analysed in relation to location, strain, climatic conditions, production system (cage, free range), housing system (sawtooth shed, hi-rise, controlled environment shed), diet (mash/pellet; home mix/commercial; enzymes/no enzymes; cereal on which diet based, for both the longitudinal and cross-sectional studies.




Flock Health and Disease Management Project Title

Trialing emergency animal disease arrangements in the Australian egg industry (EIDF)

RIRDC Project No:

AEI-10A

Start Date: 01/11/00 Finish Date: 30/09/01 Researcher: Mr Malcolm Peacock Organisation: Australian Egg Industry Association (AEIA)


Phone: (02) 9570 9222 Fax: (02) 9570 9763 Email: enquiries @aeia.org

Objectives

• To complete a trial of new emergency animal disease arrangements • To evaluate the role and effectiveness of AEIA in the management and

operation of an emergency animal disease response • To facilitate an earlier return of Australia's poultry health status to one which is

recognised internationally as being free of virulent Newcastle disease virus

Current Progress

Background Virulent Newcastle disease virus has been found on five commercial layer farms in NSW since the eradication programs undertaken on Mangrove Mountain and in Sydney in 1999. One farm is located in Tamworth (Tamworth Designated Risk Area) and four in Western Sydney (Cumberland Designated Risk Area). Within the Cumberland Designated Risk Area (DRA) there is also a "Dangerous Contact" farm and a "Suspect" farm. Government cost sharing arrangements used to fund eradication in 1999 were not activated in 2000 and consequently these farms remained infected. New cost sharing arrangements involving industry and the Commonwealth, State and Territories Governments are currently under negotiation. This project was designed to use the current situation to trial the proposed new cost sharing arrangements and to evaluate the roles of industry and government under these arrangements. Management of the Project It was decided by the National Newcastle Disease Management Committee (chaired by the Secretary of Agriculture, Forestry and Fisheries, Australia) to set up a Newcastle Disease Implementation Committee to manage these new arrangements, to be chaired by the Project Controller, Malcolm Peacock (Industry) and to include representatives of the Standing Committee on Agriculture and Resource Management, NSW Agriculture, the chicken meat and egg industries and the Industry Site Controllers for each DRA. All technical matters are referred to the Newcastle Disease Technical Committee,


which is made up of the Chief Veterinary Officers for the Commonwealth and the States and the technical advisers for the chicken meat and egg industries. NSW Agriculture and the Egg Industry are responsible for financial management of the project with all expenditure requiring approval of both parties. Operational Tasks Finalise depopulation and clean-up arrangements for the infected farms in accordance with AUSVETPLAN and the established Standard Operating Procedures. Develop replacement programs and egg supply arrangements for the infected farms. Appoint Industry Site Controllers for the Tamworth and Cumberland DRAs to monitor and complete effective audits of depopulation and clean-up procedures in conjunction with NSW Agriculture. Appoint an independent valuer for compensation claims. Provide ongoing reports to the Consultative Committee on Emergency Animal Diseases (CCEAD), RIRDC and Industry. Progress All of the infected farms have voluntarily been depopulated and cleaned-up. Owners of infected farms have received compensation. Final confirmation of satisfactory clean up through monitoring of sentinel birds is expected in early June 2001. Sentinel birds have been placed on the Dangerous Contact Farm and the Suspect Farm and test results should also be completed in early June 2001. A program of surveillance will continue for 6 months in the DRAs to substantiate freedom from virulent Newcastle Disease. Arrangements are being made to appoint a consultant to evaluate the management and operation of this emergency disease response and to prepare a full report on the exercise for RIRDC and CCEAD.


Project Title

Detection of virulent strains of Newcastle disease virus in chickens previously infected with Australian strains of the virus

RIRDC Project No:

CSA-1J

Start Date: 01/07/97 Finish Date: 30/06/01 Researcher: Dr Harvey Westbury Organisation: CSIRO Livestock Industries



Objective

• To increase the speed, efficiency and confidence of detection of virulent strains

of Newcastle disease virus (NDV) in flocks previously infected with endemic, lentogenic strains of the virus.

Current Progress

A conventional virus capture ELISA (vELISA) for Newcastle disease virus was developed using a specific polyclonal antiserum as the capture antibody and a mixture of three monoclonal antibodies as the detection system. Positive/negative levels in the test were assessed and the best target tissues in chickens infected with virulent strains of NDV determined. These were found to be bone marrow, spleen and kidney, though regular detection was also obtained from blood and lung samples. The vELISA was also able to detect virulent ND virus in the tissues of chickens immune to the disease following earlier immunisation with ND vaccine. Immune chickens were challenged with either the virulent Herts, Texas or Australian virulent ND virus strains. These virus strains could be detected in the tissues of immune chickens for up to 21 days after challenge, with most detection occurring between 4 and 10 days after challenge. The test was used on tissues of chickens naturally infected with virulent ND virus that were collected during the Australian epidemic. Results obtained from testing these samples were not as clear-cut as those obtained with tissues from experimentally infected chickens. The reasons for this difference are being examined.


Project Title

Postgraduate scholarship – Ms Louise Hilton: Therapeutic applications of cytokines in poultry

RIRDC Project No.:

CSA-10J

Start Date: 06/06/99 Finish Date: 05/06/02 Researcher: Ms Louise Hilton and Dr John Lowenthal Organisation: CSIRO Livestock Industries



Objective

• To enhance disease resistance and vaccine efficacy in poultry by

administration of cytokine therapy.

Current Progress

Cytokines are proteins that are naturally produced by the body’s immune system immediately following an infection. Cytokines protect against disease by controlling the immune response to infection or vaccination and therefore represent excellent, naturally occurring therapeutics. Previous poultry trials conducted by this research team have shown that treatment with a cytokine, interferon gamma, led to improvements in health and resulted in improved weight gains of the order of five to ten per cent. This cytokine also helped protect birds against coccidiosis infection. It is hoped that a range of therapeutics, using various types of cytokines, can be developed to provide producers with an alternative to in-feed antibiotics. This project has focused on the cytokine, chicken interleukin-2 (ChIL-2). Biologically active recombinant ChIL-2 was produced and its effects on the chicken immune system assessed. Tools were developed to investigate the activities of ChIL-2, including monoclonal antibodies which are used in an ELISA for measuring the level of this cytokine. ChIL-2 treatment of birds resulted in proliferation of both CD4+ and CD8+ populations of T cells. This effect indicated that ChIL-2 may be able to enhance cell mediated immunity, resulting in greater protection against a variety of viral and parasitic diseases. Currently, commercial trials are underway to study the ability of ChIL-2 to enhance vaccine efficacy in protection against infectious bronchitis virus and coccidiosis.



Project Title

Molecular epidemiology of Newcastle disease virus in Australia

RIRDC Project No:

CSA-11J

Start Date: 01/06/99 Finish Date: 30/06/02 Researcher: Dr Allan Gould Organisation: CSIRO Livestock Industries



Objectives

• To develop a better understanding of the epidemiology of Newcastle disease

virus (NDV) within Australia. • To develop a better understanding of the mutation rates as well as disease

potential of Australian NDV isolates at the molecular level.

Current Progress

Investigation of isolates associated with the outbreaks of Newcastle disease in NSW between 1998 and 2000 have shown that the progenitor virus for the outbreak arose, not from and exotic incursion of virulent virus, but from mutation of an avirulent, Australian precursor virus. Gene sequence and phylogenetic analysis of Australian NDVs isolated from 1932 to 2001 has identified one locus as a predictor of viral lineage and the likely ancestor virus for the progenitor virus has been identified. Viruses involved in the summer respiratory disease syndrome have been identified as belonging to two separate clades and virulent virus has been isolated from both clades. The entire genomes of eight viruses from these outbreaks have been sequenced and the genetic stability of these isolates determined. Selection pressure has been shown to be greatest for the haemagglutinin-neuraminidase, matrix and phosphoprotein genes. Analysis of the quasi-species or individual gene sequences present (at a low frequency) in field isolates has shown that virulent viruses were present in a background of avirulent progenitor virus. Variants in the fusion protein cleavage site (which determines virus virulence) have been isolated and characterised. Quasi-species analysis of virulent and avirulent plaque purified viruses have shown that two to three passages in vivo or in vitro were needed to attain the same genetic diversity as that identified in field isolates.

Studies of mechanisms for the natural selection of virulent viruses from field isolates have commenced.


Project Title

Postgraduate Scholarship - Jacqueline Kattenbelt: Mapping of structure-function relationships of Newcastle Disease (ND) using reverse genetics

RIRDC Project No:

CSA-13J

Start Date: 01/07/00 Finish Date: 30/06/03 Researcher: Ms Jacqueline Kattenbelt and Dr Allan Gould Organisation: CSIRO Livestock Industries



Objectives

• To develop a viable DNA construct containing Newcastle disease virus (NDV)

genome and separate clones of NDV nucleocapsid, protein (N), phosphoprotein (P) and polymerase (L).

• To establish a reverse genetics system to allow recovery of recombinant NDV. • To recover NDV mutants with substitutions in precise sites within the matrix

protein to give strictly defined molecular mutants. • To map structure-function relationships of viral proteins within infected cells

and to study viral morphogenesis, virulence factor and tissue trophism determinants essential for NDV replication and infection using electron microscopy.

Current Progress

Long overlapping fragments from the Peats Ridge NDV (precursor of the virulent virus which was responsible for outbreaks of Newcastle disease (ND) in Australia) have been generated and clones confirmed by polymerase chain reaction (PCR) screening and sequencing. These fragments are currently being joined at shared restriction sites to form a contiguous DNA fragment. Separate clones of nucleocapsid protein (NP), phosphoprotein (P) and RNA directed RNA polymerase (L) have been generated and cloned into expression plasmids containing an internal ribosomal entry site allowing cap independent translation. The matrix (M) gene is currently in the process of being manipulated to insert unique restriction sites to enable the gene to be cut out and a modified M re-inserted.




Project Title

Diagnostic tools for differentiation of vvIBDV and characterisation of Australian strains

RIRDC Project No:

CSA-15J

Start Date: 01/07/00 Finish Date: 30/06/03 Researcher: Dr Jagoda Ignjatovic Organisation: CSIRO Livestock Industries

Private Bag No 24 GEELONG VIC 3220


Objectives

• To introduce and compare RFLP methods developed by other research groups. • To test the specificity of Crab-88 and Crab-cos recombinant antibodies for

very virulent infectious bursal disease virus (vvIBDV) • To compare soluble, purified & phage expressed antibody for their suitability

as ELISA reagents.

Current Progress

The RFLP method was introduced and evaluated using the following overseas strains available at AAHL: classical 52/70, 1/68 and APHIS, variants E and GLS, and vvIBDV strains CS88 and Tasik. Restriction profiles generated for each virus using restriction enzymes BstN1, MboI and SspI were identical to those produced by the Jackwood and Sommer researchers who developed this method. Australian strains used in the study of Jackwood were obtained from the same source. The RFLP profile for these as well as for all other Australian strains was obtained. None of Australian strain contained an SspI site characteristic of vvIBDV strains. The specificity of the antibody Crab88 for vvIBDV strains was tested in two overseas laboratories, France and the USA. Crab88 reacted only with vvIBDV strains and did not react with any of the other classical or variant strains, including ten vaccine strains. This indicates that Crab88 is specific for all vvIBDV strains. Two other recombinant antibodies reacted with all IBDV strains, including the USA variants. Crab88 expressed as phage and soluble antibody was evaluated for use in ELISA. Higher absorbances were obtained with phage than soluble antibodies. Reagents to produce phage antibodies were cheaper and ELISA based on phage antibodies was faster and simpler.


Project Title

Attenuation and characterisation of chicken Eimeria for live vaccines

RIRDC Project No:

DAQ-259J

Start Date: 01/11/99 Finish Date: 31/12/02 Researcher: Dr Wayne Jorgensen Organisation: Department of Primary Industries (Qld)



Objectives

• To develop attenuated lines of E. mitis, E. brunetti and E. praecox for

incorporation in an efficacious, live vaccine, protective against all seven species of Eimeria in Australian chickens.

• To develop a trial technique to evaluate coccidiostat resistance.

Current Progress

The sensitivity of two strains of Eimeria mitis (Jorgensen and Kelly) to the coccidiostats Toltrazuril, Amprolium or Sulphaquinoxaline has been evaluated in trials. Both strains were sensitive to Toltrazuril and Amprolium. Each has since undergone selection for precocious development by passage (Jorgensen strain: 9 passages with a 20 hour drop in prepatent period; Kelly strain: 13 passages with a 14 hour drop in prepatent period). The Jorgensen strain was chosen for further characterisation studies and has been successfully tested for virulence, reproductive potential and protection against homologous challenge. A trial to assess the strain’s protection against heterologous challenge was aborted early and has now been repeated. The results of the new trial are now awaiting statistical analysis. Both strains of E. mitis have passed quality control and DNA testing for purity. The sensitivity of two strains of Eimeria brunetti (Monarto and Bowden) to the coccidiostats Toltrazuril, Amprolium or Sulphaquinoxaline has been evaluated in trials. Both strains were sensitive to Toltrazuril and Amprolium and are currently underging selection for precocious development by passage. The sensitivity of one strain of Eimeria praecox (Kelly) to the coccidiostats Toltrazuril, Amprolium or Sulphaquinoxaline has been evaluated in a trial. One potentially coccidiostat resistant isolate has been collected at Pitsworth, South East Queensland and cryopreserved for future study.


Project Title

Studies of cloacal haemorrhage, egg peritonitis, vent trauma and beak trimming in the laying hen

RIRDC Project No:

DAV-170A

Start Date: 01/07/99 Finish Date: 31/07/01 Researcher: Dr Greg Parkinson Organisation: Department of Natural Resources & Environment (Vic)

Victorian Institute of Animal Science 475-485 Mickleham Road ATTWOOD VIC 3049


Objectives

• To develop management strategies that ameliorates the incidence of egg

peritonitis, prolapse and picking behaviours (vent trauma and cannibalism) in the laying hen.

• To reduce flock mortalities from approximately 10% to 7% by strategic control of prolapse, egg peritonitis and vent trauma (picking).

• To stimulate the adoption of improved beak trimming practice.

Current Progress

Laboratory models have indicated that oviduct haemorrhage and/or picking behaviours in commercial layers are not uniformly distributed across the life of the laying flock. These problems are accentuated at stages that correspond to periods of high metabolic pressure (peak production and peak egg mass). Comparative studies with single-bird cage flocks indicates that approximately 50% of the cloacal haemorrhage can occur independently of picking behaviours. Furthermore, the incidence of cloacal haemorrhage appears correlated with low body weights in early lay and production of disproportionately large eggs. Birds that experience cloacal haemorrhage in early lay can continue to manifest the problem, whilst some birds repair the damaged oviduct very rapidly. Superior management of the transition from pullet to layer and more attention to body weight management will reduce the extent of cloacal haemorrhage. Two large Victorian Egg Farms with controlled environment shedding have been experimenting with non-beak trimmed flocks. In both instances annual mortality patterns have been at acceptable standards ( 5-7% ), but it is clear that a more uniform distribution of shed light intensity at 10-20 lux will lower mortality by an additional 1-2%. In the future, a combination of improved body weight management together with control over onset of sexual maturity, and uniform shed light intensities of 10-20 lux will significantly lower flock mortalities. Successful application of these approaches will facilitate the elimination of beak trimming as a routine husbandry practice.


Project Title

National NDV Survey

RIRDC Project No:

MS990-40

Start Date: 10/04/00 Finish Date: 31/07/01 Researcher: Dr Vivien Kite Organisation: Chicken Meat Program of RIRDC

PO Box 579 NORTH SYDNEY NSW 2059


Objectives

• To collect information on the type and distribution of Newcastle disease

viruses in Australian poultry flocks. • To collect information on the sero-prevalaence of NDV positive flocks across

the Australian commercial poultry indsutries and to identify risk factors for exposure to NDVs on Australian poultry farms.

• To identify possible risk factors for exposure to NDVs on Australian poultry farms.

Current Progress

The survey was designed to collect information on the sero-revalence of NDV positive flocks across the Australian commercial poultry industries, to identify possible risk factors for exposure to NDVs, and to collect information on the type and distribution of NDVs in Australian poultry flocks. The survey sampling strategy was designed to ensure comprehensive coverage of all sectors of the Australian commercial poultry industry. A total of 754 farms, across eleven regions of Australia, were sampled as part of the survey. Four types of commercial poultry enterprise were included in the survey viz. layer farms, meat chicken farms, breeder (meat and layer) farms, and dedicated pullet rearing farms. In each region, a minimum of 25% of layer farms (50% in NSW) and 30% of meat chicken farms represented in the region were included in the survey. All currently active breeder and pullet rearing farms in each region were included in the survey, except where the birds were too young. The survey was conducted in three phases. Initially, blood samples were collected and tested to establish the serological status of farms. Tracheal and cloacal swabs were then collected from seropositive farms for attempted virus isolation. Viruses isolated were genetically characterised (‘typed’), with typing based on nucleotide sequencing of the cleavage site of the gene that codes the fusion protein of NDV. A total of 259 confirmed NDV isolates were characterised, representing farms in Queensland, NSW, Victoria, Tasmania and South Australia. The majority of these isolates have come from Victorian farms. No virulent NDV isolates were detected. No precursor-like viruses (such as Peat’s Ridge or Somersby-type variants) were detected. All viruses detected have been V4-like viruses. Some minor genetic diversity was seen in these V4-like viruses,



but all are genetically distinct from the virulent virus. A full analysis of the results of the survey is currently underway.


Project Title

Molecular diagnostic tools for wild type and vaccine strains of Marek's disease virus

RIRDC Project No:

UJC-7A

Start Date: 01/09/99 Finish Date: 30/09/02 Researcher: Dr Graham Burgess Organisation: James Cook University

Dept of Microbiology and Immunology TOWNSVILLE QLD 4811


Objectives

• To develop improved procedures, which can detect genome of Type 1 Marek's

diseases virus (MDV) in the blood of infected or vaccinated birds. Develop sensitive specific PCR procedures for detecting types 2 and 3 Marek's disease viruses.

• To identify sequences that can be used to differentiate wild type and vaccine strains of Type 1 Marek's disease virus.

• To develop and evaluate simplified sample collection techniques and indicator systems that match the requirements and capabilities of the Australian poultry industry and transfer these to relevant Australian laboratories.

Current Progress

A number of blood sample collection methods have been trialed and untreated filter paper has been eliminated as a viable media. We have successfully developed a treated paper product and used capillary tubes to separate lymphocytes for transfer to filter paper. We currently have a semi-nested PCR for type 1, single and nested sets for HVT and working primers for type 2. The type 1 primers have detected CVI 988 (Rispens) following vaccination in layer birds. HVT primers are capable of detecting down to one quarter dose in broilers as a nested PCR and full dose as a single set, both using treated filter paper media. Half dose HVT vaccination can also be detected with a single primer set and the capillary tube collection system. We can now differentiate between CVI 988 and Australian strains using the meq gene of extracted and purified DNA. Pulsed field gel electrophoresis is underway to separate viral genome in pure form for restriction digestion and cloning. This will be used for sequencing and as a plasmid DNA library for Australian strains of MDV. An ELISA format for product detection has been chosen and we are currently designing probes for all three serotypes and several optimising blocking reagents.



Project Title

Determination of the genomic sequence of Mycoplasma gallisepticum

RIRDC Project No:

UM-45J

Start Date: 01/06/99 Finish Date: 15/06/01 Researcher: Dr Glenn Browning Organisation: The University of Melbourne



Objectives

• To determine the complete genomic sequence of Mycoplasma gallisepticum. • To facilitate identification of genes which are likely to play a role in virulence. • To lay a foundation for subsequent studies to improve the performance of

mycoplasma vaccines and to improve diagnosis of mycoplasmosis.

Current Progress

A library of clones has been constructed for sequencing. Draft coverage of the genome, with an estimated three fold redundancy, has been obtained to date. Although this data has been useful it is not in a form that can be dispersed yet, as it comprises numerous small contigs. A few gaps will remain to be closed after the completion of the random cloning phase and that is expected to take another several months to complete. The annotation will begin after that time.


Project Title

Investigating sanitation of surface water for poultry using chlorine - IBDV models

RIRDC Project No:

UM-51A

Start Date: 01/07/00 Finish Date: 31/07/01 Researcher: Dr Trevor Bagust Organisation: The University of Melbourne

Faculty of Veterinary Science, Pre-Clinical Centre Cnr Park Drive & Flemington Road PARKVILLE VIC 3052


Objectives

• To increase biosecurity for poultry production sites, which use surface water. • To gather objective pilot-scale scientific data about procedures required for the

most economical yet effective inactivation of important poultry viruses in surface water of varying qualities.

• To scale-up the above pilot system as a step towards enabling cost-effective on farm use.

Current Progress

Ensuring that the water supplies used in poultry production are adequately sanitised is central to site biosecurity. Surface waters, which may include dams, lakes, rivers or creeks are widely used in rural Australia as water sources. However, as these are also open to access by waterfowl, there is significant risk for potential contamination with poultry pathogens (disease-causing micro-organisms). Of continuing concern at present are the viruses of Newcastle Disease, avian influenza and egg-drop syndrome adenovirus, all of which may spread with migratory ducks. A further threat, which has emerged over the last decade, are very virulent pathotypes of infectious bursal disease virus (IBDV). Now spread throughout Asia, these are of much greater pathogenicity for chickens than our endemic IBDV strains .The scientific literature has no data on the effectiveness of commonly-used water treatments e.g chlorine or its derivatives, for these poultry viruses. Research studies being progressed here during the last year have determined that: (1) Newcastle Disease virus (NDV) diluted to 100-1000 infectious units per ml in potable drinking water, pH 7.0, can be effectively inactivated by treatment levels of 1ppm free chlorine within 1 hr, providing that this level of sanitation is maintained e.g. using a chlorine injector- batch treatment system. (2) Near-instantaneous drops in free chlorine concentrations, to sub-effective sanitation levels, have been detected to have occurred within 1 minute when attempting to sanitise water supplies containing even low levels (to be quantified) of protein. (3) Chlorine dioxide at a starting treatment concentration of 1ppm is also completely effective in inactivating NDV in drinking water, but continues to be



effective at lower residual levels than chlorine. (4) Laboratory Australian strains of IBDV have been examined for sensitivity to chlorine, using the standardised treatment and chlorine-monitoring systems developed during these studies. Preliminary results obtained with IBDV strain GT101 indicate that IBDV virus in drinking water would appear to be much more resistant to the inactivating effects of chlorine than is NDV. We are now undertaking the preparation of concentrated purified IBDV stocks, in order to enable closer quantifying of dose-inactivation of Cl for IBDV in drinking water. We will compare these with ClO2, then move to extend sanitation treatments to testing in samples of surface water.


Project Title

Control of intestinal spirochaete infections in chickens

RIRDC Project No:

UMU-23J

Start Date: 01/06/99 Finish Date: 30/08/01 Researcher: Prof David Hampson Organisation: Murdoch University

Division of Veterinary and Biomedical Sciences MURDOCH WA 6150


Objective

• To identify new and appropriate means to control infection by the intestinal

spirochaetes, Brachyspira intermedia and Brachyspira pilosicoli, recently recognised and common pathogens causing significant economic loss in Australia layer and broiler breeder flocks.

Current Progress

Studies have been conducted to determine in vitro antimicrobial drug sensitivities of spirochaete isolates from Australian chickens, and to test in vivo antimicrobial activity in experimentally infected layer and broiler breeder birds. Testing of 66 spirochaete strains revealed a similar spectrum of sensitivity to antimicrobial drugs as porcine spirochaetes. Resistance was seen against a number of drugs, with the least resistance recorded for tiamulin. In vivo, tiamulin at 25mg/kg body weight cleared experimental infection with Brachyspira intermedia in layer hens. Birds in an infected room, however, became re-infected after treatment ceased. This suggests that it may be better to use continual low level therapy to control these infections, or to use therapeutic levels of drugs combined with a thorough environmental cleaning program. Continued in-feed supplementation with zinc bacitracin at 100 ppm inhibited proliferation of B. intermedia in experimentally infected birds, whilst at 50 ppm zinc bacitracin encouraged proliferation of Brachyspira pilosicoli. It is uncertain whether these differences are due to the different spirochaete species investigated, or to the dose rates of zinc bacitracin used. Overall, these conflicting results suggest that there are complex interactions between the intestinal microflora and pathogenic intestinal spirochaete species, and that control will require careful monitoring.


Project Title

Effects of diet composition, gut microbial status and feed forms on cannibalism in layers

RIRDC Project No:

UNE-72A

Start Date: 01/07/99 Finish Date: 01/11/02 Researcher: Dr Mingan Choct Organisation: University of New England



Objectives

• To examine the interaction between diet composition and the incidence of

cannibalism • To investigate the effect of the gut microbial status on cannibalism in layers

Current Progress

A total of four experiments have been conducted to examine the effects of rearing conditions (low light and natural light), beak-trimming and diet composition on the onset of cannibalism in ISA Brown layers. Rearing conditions had no effect, whereas beak-trimming had a profound effect, with cannibalism occurring predominantly in untrimmed birds. In conventionally-housed, untrimmed birds, feeding high fibre diets (e.g., millrun, rice hulls) was not only effective in preventing the onset of cannibalism but also in alleviating cannibalism mortality in flocks already having problems. The mechanism of this action is not known. We have found that digesta transit rate is faster in birds fed high fibre diets, which coincides with a lower gut viscosity, a reduced incidence of pecking, and an increased frequency of feeding. Dietary energy density does not appear to be a cause because diluting the diet with sand had no preventative effect. However, diet form is important, as mash diets are more preventative than pellets, in particular when the diet has a low fibre content. Two more experiments are currently underway to investigate (1) whether these findings are repeatable, and (2) the effect of “behavioural conditioning” during rearing on cannibalism during lay in a barn system.


Project Title

Optimising infectious bronchitis vaccination of laying hens for maximum egg shell quality

RIRDC Project No:

UNE-76A




Objective

• To provide to the Australian Egg Industry recommendations concerning best

practice in infectious bronchitis vaccination protocols

Current Progress

A preliminary study was conducted to assess the age at first vaccination (day-old or two weeks of age) and the route of vaccination using the VicS strain infectious bronchitis (IB) virus vaccine. The negative control group remained free from IB whereas the antibody titres were similar for all vaccinated birds. The first trial of the main project commenced in October 2000 using Isa Brown laying hens. All birds were vaccinated initially at day-old and again at 4 weeks, except for the control group, using either VicS or A3 strains of vaccine and delivering the vaccine via one of three routes: eye drop, coarse spray or in water. There is no evidence that the A3 vaccine virus is too harsh for day-old birds. All birds, including the controls, were vaccinated using VicS by eye drop at 12 weeks. Half of the birds (Poultry Shed C) will receive no further vaccination for IB whereas the other half (Poultry Shed B) are being revaccinated every eight weeks. Egg production, body weight, feed intake, blood electrolytes, IB antibody titres, manure moisture, egg and egg shell quality are being monitored at regular intervals. All mortalities are being autopsied. Birds are currently 30 weeks of age.




Project Title

Enhancing mucosal immunity in chickens by novel in-ovo and postnatal vaccination techniques

RIRDC Project No:

US-72J

Start Date: 01/01/99 Finish Date: 31/12/01 Researcher: Prof Alan Husband Organisation: The University of Sydney

Dept of Veterinary Anatomy and Pathology UNIVERSITY OF SYDNEY NSW 2006


Objective

• To induce long-term immunoenhancement in chickens following early priming

of the avian immune system via in-ovo immunisation through: − development of naked DNA or recombinant constructs of antigen and/or

cytokines; − evaluation of delivery vehicles such as liposomes or biodegradable

microspheres; − assessment of immunoregulators for non-specific upregulation of the

immune system; and − evaluation of immunisation protocols for enhancing immune responses to

routine vaccinations and providing protection from disease challenges such as Salmonella Typhimurium.

Current Progress

Initial contact with and colonisation of the intestinal mucosa by pathogens is impeded by IgA antibody located at the intestinal surface. Appropriate vaccination procedures will stimulate intestinal IgA production, protecting the host from these pathogens. Such IgA antibody production can be increased through the use of immunoenhancers. Studies undertaken have investigated the immunoenhancing potential of vitamin E (VE) or the cytokine interleukin-6 (IL-6), (a communicator of the immune system, whose mammalian counterpart increases IgA production) to increase IgA antibody levels following vaccination in chickens. Dietary VE supplementation, from day old, increased antigen-specific intestinal IgA antibody titres following immunisation with either tetanus toxoid or whole killed Salmonella Typhimurium. At some dose rates, in-ovo delivery of VE with killed S. Typhimurium antigen elicited an increase (not statistically significant) in anti-S. Typhimurium IgA antibody levels post-hatch. Repeated oral delivery of IL-6 to chickens immunised with either tetanus toxoid or whole killed S. Typhimurium increased anti-antigen IgA at the intestinal surface. A live S. Typhimurium challenge model is being established to examine the ability of IL-6 induced increases in IgA antibody following immunisation, to protect chickens from a challenge of S. Typhimurium.


Feed Availability and Nutrition Project Title

Characterisation of canola meal and cottonseed meal at practical inclusion levels for use in broiler and layer diets

RIRDC Project No:

DAQ-264J

Start Date: 01/07/99 Finish Date: 31/12/01 Researcher: Dr Rider A Perez-Maldonado Organisation: Department of Primary Industries (Qld)

Queensland Poultry Research and Development Centre PO Box 327 CLEVELAND QLD 4163


Objectives

• To measure the variability of glucosinolates, sinapines, condensed tannins

(CT), total phenolics (TP), sulphur and phytic acid levels in canola meal (CM) and CT, TP and free and bound gossypol levels in cottonseed meal (CSM). Pesticide residue levels in CSM; AME, amino acid and proximate composition of CM and CSM will be determined in samples from major processing sites, three times a year.

• To evaluate the ratio of iron to free gossypol in CSM to optimise production in both broilers and layers.

• To determine the upper limits of inclusion of both CM and CSM separately and in combination in broiler and layer diets.

• To make recommendations to the poultry industries on the nutritional value of both CM and CSM when included in least-cost poultry diets at levels close to their upper limit.

Current Progress

During 2000/01 several experiments were conducted to investigate the effects of CSM or CM fed at levels of 10, 20, 30 or 40% of the diet to broilers and laying hens. At 21 d chicks fed CSM showed a reduced feed intake (FI) from the 20% level. Liveweight gain (LWG) was also reduced at 20 and 30% CSM in the diet but not at the 10 and 40% levels. The feed conversion ratio (FCR) was only affected at the 20% level, where it was poorer. After 37 d LWG and FCR were not affected by any level of CSM, indicating that older birds were capable of overcoming any negative effect of CSM observed in younger birds. The results for CM indicated that after 21 d chicks fed Pinjarra CM had improved FCR at all levels, with a reduced FI but a good LWG at all but the 40% level. After 37 d a similar pattern of bird performance was observed. After 21d chicks fed Numurkah CM had a reduced FI from the 20% level but there was no effect on FCR and LWG even at the 30% level. After 37 days birds showed a similar FI and LWG response but with significantly improved FCR at all levels. After 21 d



Newcastle CM resulted in a poorer FCR from the 30% level but a good LWG and FI was observed at all levels except the 40% level. However, after 37d the birds overall FI and LWG was reduced from the 20 and 30% level, respectively, without affecting the overall FCR. After 21 d Melbourne CM resulted in a reduction in LWG and FI from the 20 and 30 % levels, respectively. However, after 37d an improved FCR was observed at all levels, even though FI and LWG were reduced from 20% level. Oil processing conditions and the presence of anti-nutritional factors may have influenced the overall performance in these meals. Although these results demonstrate that substantial amounts of CSM and CM can be used in broiler diets, more detailed studies are being undertaken to confirm the above findings. In the layer experiments, good performance was obtained when feeding CSM or different sources of CM at levels of 10, 15, or 20% of the diet. However, preliminary observations made to evaluate fresh and stored eggs derived from the above experiments indicated that an abnormal odour (fishy taint) was detected in raw eggs derived from brown layers on all CM diets. Due to the importance of this observation, further work is being undertaken using an expert sensory panel to evaluate eggs derived from hens fed CM and CSM diets.


Project Title

Inclusion of data for additional livestock species in the Australasian Livestock Feed Ingredient (ALFI) database

RIRDC Project No:

GRD-2J

Start Date: 01/04/99 Finish Date: 30/06/01 Researcher: Dr Robert van Barneveld Organisation: Grains Research and Development Corporation

C/- Barneveld Nutrition Pty Ltd PO Box 42 LYNDOCH SA 5351


Objective

• To improve knowledge of the nutritional value of feed grains and the

efficiency of use of these grains by the egg and chicken meat industries through development of a commercial version of the Australasian Livestock Feed Ingredient (ALFI) database containing data for pigs, poultry (layers and broilers) and aquaculture species.

Current Progress

The Australasian Livestock Feed Ingredient (ALFI) database now contains more than 22,332 sample entries on the chemical composition of feed ingredients and the nutritional value of these ingredients for pigs, poultry (layers and broilers) and aquaculture species. ALFI also incorporates a vast range of information contained in recent literature and other relevant databases. The re-programmed version of ALFI has been tested by the major stakeholders and the programming of the database is now complete. A business plan has been prepared for commercialisation of the ALFI database based on distribution to end-users as a subscription service via the internet or as a CD-ROM. Final negotiations with stakeholders on commercialisation are under way.



Project Title

Premium Grains for Livestock Program (stage 2)

RIRDC Project No:

GRD-3J

Start Date: 01/07/00 Finish Date: 30/06/03 Researcher: Dr John Black Organisation: John L Black Consulting



Objective

• This project is the second stage of a major research program for improving

feed grains quality and marketing that has been negotiated in response to identified industry needs. It has five integrated component projects:

1) coordination 2) production, storage and distribution of grain samples 3) rapid and objective analytical tests for assessing feed grains quality 4) enhancing grain nutritional value through breeding and processing and 5) modelling feed grain quality.

Current Progress

Several hypotheses about the factors determining the nutritional value of cereal grains for ruminants, pigs and poultry were established in the predecessor Project GRD-1J. The relative proportion of the main chemical components of a grain is the major determinant of nutritional value for all classes of livestock. However, other grain characteristics can significantly affect energy availability and these differ between ruminants, pigs and poultry. Prediction of AME in poultry based on an assumed constant digestion of individual gross chemical components of grains show close agreement with observed values for sorghum and oats, but not for wheat or barley. The accuracy of predictions for triticale was intermediate between sorghum and barley. Further analyses showed that some of the difference between predicted and observed AME values could be explained by the viscosity of ileal digesta. However, other factors such as grain hardness and hydration capacity appear to be associated with the variation in AME between grains. In addition, starch characteristics such as granule size and distribution, amylose:amylopectin ratio and gelatinisation temperature may influence the extent of starch digestion in the small intestines of poultry and therefore energy availability. These hypotheses will be tested further in poultry over the coming year.


Project Title

Hind gut function in laying hens

RIRDC Project No:

UNC-12A

Start Date: 20/09/99 Finish Date: 21/09/01 Researcher: Dr Robert Taylor Organisation: The University of Newcastle

Nutrition & Dietetics Level 3 Medical Sciences Building CALLAGHAN NSW 2308


Objective

• The project is designed to provide evidence of hind gut acidosis in laying birds

and to determine strategies to reduce or eliminate this condition.

Current Progress

The possibility of acidosis in the digestive tract of the chicken, as found in other livestock species, was discounted in a recent publication. The consequences for the bird productivity, efficiency and/or health, if acidosis was produced by rapid fermentation of carbohydrate, would be profound. The project aimed to determine if a pH reduction in the “hindgut” of the bird could be produced. Experiments were based on wheat, rice, barley and sorghum diets in birds raised from day-old and fed commercial starter, grower and layer diets. Birds were used at different points in the production cycle from young growers to layers at peak production. The cereals were incorporated in the diets in various forms (standard grind versus whole grain incorporation into crumbles); commercial feed enzymes were used and diet presentation was varied by feeding “meals” and wet mash. Initial results suggest that digesta pH in the lower segments of the digestive tract can attain low pH within the first 48 h of changing feed type and processing. Additional work with broiler birds confirmed that gut pH can rapidly decrease and may attain levels consistent with those in animals suffering acidosis. The mechanism is unclear. Further, it appears that the quantity of feed is important at the particular stage of life i.e. greater feed intake consistent with mature body size and production level influences the response to diet change. The weather conditions at the time of feed change may interact with the cereal type fed change to exacerbate the condition. Specifically, in hot weather, increased water intake may induce more rapid throughput of feed leading to greater substrate for fermentation in the lower gut. Short chain fatty acid production in the caeca, ileum and other gut segments indicates that fermentation may occur high in the intestinal tract and may result in ratios of acetate:propionate:butyrate that are different in commercial poultry than other classes of animals. The presence of iso- forms of SCFA indicated that protein fermentation may occur through the intestinal tract. Analysis of samples and data is continuing.



Project Title

Effects of commercial feed enzymes in wheat-based diets on egg and egg shell quality in imported strains of laying hen

RIRDC Project No:

UNE-77A




Objective

• To provide, to the Australian Egg Industry, recommendations concerning the

advantages and disadvantages of the use of commercial enzyme preparations in layer feed.

Current Progress

Isa Brown pullets, 16 weeks of age, were transferred to the university farm in November 2000. Birds received prelayer diet initially, layer diet at 19 weeks and one of ten experimental diets at 25 weeks of age. Diets are based on one of two types of wheat: “standard” or “pinched” wheat. For each wheat type, diets contained either no enzyme (control) or one of four different types of commercial feed enzyme preparations. Eggs were collected for analysis at 27 weeks and 30 weeks, then at 5 weekly intervals (ongoing). Apparent Metabolisable Energy (AME), feed intake, manure moisture and body weight were measured at 30 weeks and then at 5 weekly intervals (ongoing). Analysis of the two wheats showed that they were similar for soluble, insoluble and total non-starch polysaccharides, although the “pinched” wheat had a higher protein level. At 35 weeks of age, the AME value for the control birds was lower for the pinched wheat. However the addition of feed enzymes increased the AME value of the pinched wheat to that of the standard wheat. Egg and egg shell quality appear to be influenced by both the wheat type and the addition of enzymes.


Project Title

Evaluation of Lathyrus cicera as a feed ingredient for layers

RIRDC Project No:

UWA-61A

Start Date: 01/07/00 Finish Date: 31/12/01 Researcher: Dr Colin Hanbury Organisation: University of Western Australia

Centre for Legumes in Mediterranean Agriculture (CLIMA) NEDLANDS WA 6907


Objective

• To establish Lathyrus cicera cultivar Chalus as a low cost grain legume of

sufficient quality to be included in layer hen rations. This will be an evaluation of layer performance, visual observation, egg quality and incidence of soiled eggs at inclusion rates up to 30% Lathyrus.

Current Progress

The project is divided into 3 parts, an initial long term trial, a large layer production trial and a final long term study. The initial long term feeding trial was completed in December 2000 and was designed to look for residues of ODAP (a toxin) in eggs and bird tissues when fed cv Chalus (a cultivar known to have very low levels of ODAP). It was demonstrated that there was no carry over of ODAP into eggs or tissue. Despite not being a production study the initial trial indicated that egg production and feed intake results were typical for birds of this breed and age and that 15% cv Chalus and possibly up to 30% could be included in the diet without detriment to performance. Following the successful outcome of the initial trial a larger trial of layer production was commenced in May 2001, the purpose being to evaluate layer performance and egg quality characteristics. To date there have been no significant differences in laying rates between controls and the range of inclusion levels of cv. Chalus (5, 10, 15, 25, 30%) in the diet. The complete data for this trial will not be available for analysis until July 2001.




Husbandry and Welfare Project Title

Should claw abrasives be used in cages in Australia?

RIRDC Project No:

SAR-34A

Start Date: 01/01/01 Finish Date: 31/01/02 Researcher: Dr Phil Glatz Organisation: South Australian Research and Development Institute

PIRSA/SARDI Research Funds Coordinator LSA Building, PPPI Roseworthy Campus ROSEWORTHY SA 5371


Objectives

• To determine the effect of abrasive strips and abrasive paint in layer cages on

claw length and sharpness, foot condition, feather cover, body scratches and mortality of a Hyline Brown strain.

• To determine if claw abrasives should be used in the Australian egg industry.

Current Progress

A trial is being conducted in the layer unit of the PPPI at Roseworthy Campus to determine the effect of abrasive strips and abrasive paint in layers cages on claw length and claw sharpness, foot condition, feather cover, body scratches and mortality of Hyline Brown hens. A previous study with Hyline Gold hens found that hen mortality from prolapse and cannibalism was significantly higher in cages fitted with abrasives. The current trial has been in progress for 3 months, without any signs of mortality from cannibalism or prolapse. The foot and body condition measurements were made on the birds prior to exposure to the abrasives. Average claw length at housing was 18.4 mm, which was similar to the claw length recorded for Hyline Gold hens (18.7 mm) in the earlier project. The body condition measurements will be repeated at the end of the trial in December 2001.


Project Title

Beak trimming accreditation

RIRDC Project No:

SAR-35A


Livestock Systems Alliance Building Roseworthy Campus ROSEWORTHY SA 5371


Objectives

• To develop quality assurance documentation to improve beak trimming. • To develop an accredited beak trimming course. • To prepare best practice beak trimming training manuals. • To assist the egg industry to achieve consistent beak trimming. • To improve public perception of beak trimming by accrediting beak trimmers.

Current Progress


mailto:[email protected];



Environmentally Sustainable Management Project Title

Reduction of dust emissions from broiler and caged layer sheds

RIRDC Project No:

SAR-33J

Start Date: 01/01/01 Finish Date: 31/12/02 Researcher: Mr Thomes Banhazi Organisation: South Australian Research and Development Institute

Pig & Poultry Insititute GPO Box 397 ADELAIDE SA 5001


Objectives

• To determine the major risk factors associated with increased concentrations of

dust within, and dust emissions from, naturally and mechanically ventilated poultry houses.

• To develop strategies that will reduce both interior dust levels and dust emissions from buildings, resulting in more sustainable housing systems for egg and broiler production in semi-urban and more densely settled areas and reduced OH&S risks for staff.

Current Progress

The project is progressing well and according to the predetermined project timetable. The location of sheds and sampling points for measuring air quality in layer and broiler sheds (dust, ammonia etc) has been determined, including the time and season of sampling. Consultation is taking place to confirm these sampling points between the research team and Dr H. Takai of Denmark, an international expert in dust related issues in animal houses. Five different types of buildings will be studied, viz. naturally and mechanically ventilated layer and broiler sheds and tunnel ventilated broiler buildings. A questionnaire on housing for industry participants to complete has been developed. The first three farm visits for monitoring air quality was completed in late May 2001. Meanwhile discussion is also taking place with a SA based engineering firm to jointly design and evaluate a simple dust trap to be used at the air outlets of mechanically ventilated poultry sheds.


Project Title

The assessment and development of best management practice techniques for Australian laying hens housed in conventional and alternative laying systems

RIRDC Project No:

UQ-93A

Start Date: 01/04/00 Finish Date: 30/06/02 Researcher: Mr Geoff Stewart Organisation: The University of Queensland

School of Animal Studies GATTON QLD 4343


Objectives

• Comparison of existing housing systems via a national survey of

commercial flocks and development of best management practices through quantitative research for free range, barn and cage production systems in relation to welfare, production, economics, air quality, egg quality, food safety, health and parasite control.

• Publish manuals for best practice in free range, barn and cage production systems in Australia.

• Provide written draft material to educate producers to enable them to achieve best practice.*

• Prepare extensive reviews on hen welfare, production, egg quality, air quality, OH & S, food safety, parasitology and economics for free range, barn and cage production systems.

• Provide material for use in educational campaigns to inform the community on the relative advantages and disadvantages of various egg production systems.*

*Note: the extension and educational components are not part of the scope of the proposed project but need to be developed as part of the outcomes for this project.

Current Progress

By RIRDC direction, the first stage of this project (The National Layer Survey) has been delayed due to political considerations associated with the invited involvement of RSPCA Australia. Furthermore, RIRDC originally directed that the second stage of this project (A quantitative comparison of conventional and alternative egg production systems) was not to commence until Part 1 is complete and has been discussed by the project’s National Advisory and Technical Committees. However, due to the delay in the commencement of the first stage of the project RIRDC has reconsidered this position and commencement of both stages of the project will be discussed at an Advisory Committee meeting planned to be held on Monday 23 April 2001. This meeting is expected to discuss the conduct of the National Layer Survey, the format of the ‘farmer attitude survey’ which will provide key indicators about the attitude of farmers and their staff to the welfare and management of their stock and the housing systems they employ. It is expected



that the National Co-ordinator will be directed to proceed immediately with Part 1 of this project following this NAC meeting. In addition to the above, it is expected that the NAC at its 23APR01 meeting will inspect facilities to be used at UQ Gatton for Part 2 of this project and will make recommendations for the acquisition of appropriate infrastructure to provide ‘worlds best practice’ facilities in which to conduct the quantitative comparative studies of the four major layer housing systems used in Australia (free range, barn, conventional cage and controlled environment cage). It is expected that it will take in the order of 6 months to order equipment, build and outfit three new free range facilities, convert the existing University broiler shed in to a three replicate barn layer shed, and refurbish part of an existing layer shed into a controlled environment shed. Given the RIRDC instruction not to proceed with the project National Survey or the subsequent Comparative Analysis of Layer housing Systems, the project National Co-ordinator has used the time since appointment to the project to collect literature review information regarding the project, and to physically prepare the University Poultry Unit facilities and site for ‘best practice’ refurbishment as to be directed by the National Advisory Committee.


Project Title

Pilot study on the use of time lapse video to study the behaviour of laying hens in conventional and modified cages

RIRDC Project No:

UQ-97A

Start Date: 01/02/01 Finish Date: 30/08/01 Researcher: Mr Geoff Stewart Organisation: School of Animal Studies

University of Queensland GATTON QLD 4343


Objectives

• To determine, via video review and analysis, the behaviour of laying hens and

their interaction with cage resources in conventional and modified cages. • To quantify problems in the use of time lapse videos as a method of studying

hen behaviour in cages. • To prepare demonstration video tapes of various behaviours to enhance the

welfare debate about cage and modified cage welfare. • To provide scientific data that can be utilised by all sectors of society to better

understand how laying hens use cage resources in conventional and modified cages.

Current Progress

As a pilot study, video footage of a light and a heavy bodyweight strain of commercial laying hen housed in conventional or modified laying cages is being analysed to compare behaviour and to investigate the use of the perch, nest and litter box supplied in the modified cage. Behaviours in each cage type and of each layer strain, including how cage spaces are used and how much different spaces are used, are being measured and the data summarised to look for trends. Individual variability is also being measured to assess the welfare impact for all individuals in the cage and to assess how many replicates would be needed for statistical analysis of this type of data. The video tapes cover a full 24-hour period. Data is being collected at a range of times throughout the day, including the first and last half hour, a half hour period every two hours during the light period and a half hour period following topping up of the food trough. Where egg laying is observed, the activity of the hen concerned is recorded for approximately the previous 2 hours and the following 10 minutes. The entire tape is viewed for use of nest and litter box by each individual. Data is collected by two methods – by scanning and by noting the time of start and stop of continuous behaviours such as sitting, and bouts of sporadic behaviours such as feather pecking. The range of behaviours being measured includes: feeding, drinking, preening, sitting, standing, position changes, non-aggressive feather pecking, aggressive pecking, other pecking, comfort movements (stretch, ruffle, wing stretch) and dust-bathing.


Training, Information and Technology Transfer Project Title

National egg industry newsletter

RIRDC Project No:

DAN-138A

Start Date: 01/07/96 Finish Date: 31/12/01 Researcher: Mr Gerry Bolla Organisation: NSW Department of Agriculture

Gosford Horticultural Research & Advisory Station Locked Bag 26 GOSFORD NSW 2250


Objective

• To maintain production of the biannual national egg industry newsletter “In an

Eggshell”. The newsletter will focus on the dissemination of latest research findings from around Australia with a special focus on improving the standard of layer housing and environmental management, bird welfare and the adoption of world's best practice in these areas.

Current Progress

The Winter, 2000 issue No. 9 was completed and mailed out in June, 2000 and included information on better housing for layers, training in the egg industry and an update on welfare issues and alternative production systems in Europe. Also included was the latest update on developments from the latest ARMCANZ meeting. The Summer, 2000 issue No. 10, completed in December, 2000, contained a number of RIRDC funded project results including the nutritional value of pearl millet, the impact of farm practices on egg shell quality, review of beak trimming methods and improving the competitiveness of the Australian egg industry. The ARMCANZ decision on the future of laying cages in Australia was also published to allow producers to familiarise themselves with this important milestone ruling to assist in future farm planning and development. The Winter, 2001 issue No. 11 is well underway and will be mailed out in June, 2001. Mailing lists have been updated following each mailout and the index of publications reduced to one page as suggested by the RIRDC Egg Program Manager.


Project Title

Video series - Workplace training for layer farm staff

RIRDC Project No:

DAN-189A

Start Date: 01/11/00 Finish Date: 31/08/02 Researcher: Mr Michael Bourke Organisation: NSW Department of Agriculture

Locked Bag 21 ORANGE NSW 2800


Objectives

• The outcome of the project will be the facilitation of workplace training for egg

farm workers that is aligned with the National Agriculture Training Package. The objectives of the project are: To develop video scripts that are aligned with National Agriculture

Training Package competencies and Murrumbidgee College of Agriculture's distance education materials on Layer Management.

To produce a series of video training materials for the egg industry.

Current Progress

A project brief has been produced during 2000-2001. This document provides a comprehensive outline for the project including the video series structure and content areas. The project brief ensures that the researchers, RIRDC and the film production company all clearly understand the project’s messages and desired outcomes. The project brief was developed with input from the RIRDC Egg Committee and egg producers. It identifies a series of four videos targeted at new employees and is linked to the industry's proposed QA program. The video titles are: • Producing Quality Eggs (an introduction to QA) • Routine Checking (stockmanship involved with observation) • Handling Birds (stockmanship involved with animal husbandry practices) • Farm Practices (other activities carried out by employees) Themes in each video will include bird welfare, QA, biosecurity and consumer concerns. These have been incorporated into a draft script for the Producing Quality Eggs video that is being refined before going to the Egg Committee for comment. Preparation of a draft script for the Routine Checking video has also commenced. Liaison with two film production companies has taken place with the intention of obtaining quotes from each to produce the videos. An audit of current training materials has commenced while ideas for the workplace training guidelines have been investigated.



Project Title

Vaccination training manual

RIRDC Project No:

SAR-36A


Livestock Systems Alliance Building Roseworthy Campus ROSEWORTHY SA 5371


Objectives

• To document best practice vaccination procedures. • To prepare vaccination training materials. • To assist the egg industry to achieve effective vaccination programs.

Current Progress


OTHER SUPPORTED ACTIVITIES 156

OTHER SUPPORTED ACTIVITIES

SCHOLARSHIPS

CHICKEN MEAT PROGRAM UNE-64A Postgraduate scholarship - Andreas Kocher: Increasing the nutritive value of grain legumes for

poultry by use of more efficacious enzyme systems Refer to report page 24 US-68A Postgraduate scholarship - Ron Newman: Manipulation of lean tissue deposition by altering the

sensitivity of tissues to the metabolic hormones Refer to report page 27 CHICKEN MEAT AND EGG PROGRAMS CSA-4J Postgraduate scholarship - Matthew Rudd: Identification of virulence determinants of infectious bursal

disease virus (IBDV) Refer to report page 3 CSA-10J Postgraduate scholarship - Ms Louise Hilton: Therapeutic applications of cytokines in poultry Refer to

report page 46 CSA-13J Postgraduate scholarship - Jacqueline Kattenbelt: Analysis of virulence determinants of Newcastle

disease virus Refer to report page 49 UTS-3J Postgraduate scholarship - David Witcombe: Production and characterisation of recombinant

antigens of Eimeria and their potential use in a maternally-delivered vaccine against poultry coccidiosis Refer to report page 18

EGG PROGRAM UM-46A Postgraduate scholarship - Ms Michelle Peters: Molecular biology of chicken anaemia virus UNS-13A Scholarship - Wei Leng (Belinda) Chung: Defined probiotic preparations for competitive exclusion of

enteropathogens from poultry

RESOURCE DEVELOPMENT

CHICKEN MEAT PROGRAM DAV-159A Animal Welfare Centre EGG PROGRAM BGC-1A Development of a national quality assurance program for the Australian egg industry MS001-02 Development of a biosecurity code for the commercial egg industry MS001-17 Computer use and training survey

OTHER SUPPORTED ACTIVITIES 157

TRAVEL/CONFERENCE/WORKSHOPS CHICKEN MEAT PROGRAM TA001-03 The Second International Conference on Air Pollution from Agricultural Operations (Air Pollution 2000),

Iowa, USA, October 2000 – Mr John Jiang CHICKEN MEAT AND EGG PROGRAMS TA001-10 Annual Conference of American Association of Avian Pathology, Utah, USA, 12-17 July 2000 - Dr Peter

Kirkland TA001-20 2001 Australian Poultry Science Symposium, Sydney, Feb 2001 TA001-24 Australian Veterinarian Poultry Association (AVPA) Scientific Meeting, Attwood, 15-16 November 2000 -

Dr Mike Alcorn TA001-30 Australian Veterinary Poultry Association Meeting, Sydney, February 2001- Dr M Folden Jansen TA001-46 7th International Poultry Welfare Symposium, Switzerland, Aug 2001 - Prof John Barnett TA001-47 2nd International Symposium on IBD and Chicken Anaemia, Germany, June 2001 - Mr Matthew Rudd TA001-48 Visit to Australia, August 2001 - Dr Daniel Todd EGG PROGRAM MS001-59 Meeting costs for the Layer Welfare Research Management Group TA001-23 International Egg Commission Annual Production and Marketing Conference, USA, September 2000 -

Mr Hugh McMaster (EIDF) TA001-26 EUROTIER International Exhibition for Livestock and Poultry Production, Germany, November 2000 -

Mr Andrew Almond TA001-28 Australian Poultry Science Symposium, Sydney, 7-9 February 2001 – Dr Phil Glatz TA001-32 6th European Symposium on Poultry Welfare, Switzerland, September 2000 – Dr Phil Glatz TA001-54 Visit to Australia, May 2000 - Professor Jorg Hartung TA001-59 6th European Symposium on Poultry Welfare, Switzerland, Sept 2001 - Mr Peter Bell TA001-60 Visit to Australia, August 2001 - Mr Andrew Joret WS990-20 Benchmarking strategy workshop WS001-08 Industry Databases Workshop

RESEARCH TO BE SUPPORTED IN 2001/2002

158



Flock Health Project Title Project No Principal Investigator Organisation Phone No Efficacy trials of a maternally-delivered recombinant vaccine against coccidiosis CME01-05J Dr Nicholas Smith University of Technology, Sydney (02) 9514 4013 Postgraduate scholarship - Ms Louise Hilton: Therapeutic applications of cytokines in poultry CSA-10J Dr John Lowenthal CSIRO Livestock Industries (03) 5227 5759 Molecular epidemiology of Newcastle disease virus in Australia CSA-11J Dr Allan Gould CSIRO Livestock Industries (03) 5227 5119 Biological control of necrotic enteritis in meat chickens CSA-12A Dr Robert Moore CSIRO Livestock Industries (03) 5227 5760 Postgradute Scholarship - Jacqueline Kattenbelt: Analysis of virulence determinants of Newcastle disease virus

CSA-13J Dr Allan Gould CSIRO Livestock Industries (03) 5227 5119

Diagnostic tools for differentiation of vvIBDV and characterisation of Australian strains CSA-15J Dr Jagoda Ignjatovic CSIRO Livestock Industries (03) 5227 5769 Evaluation of fowlpox (FPV) strains free of reticuloendotheliosis virus (REV) as vaccines for use in Australian poultry flocks

CSA-16A Dr David Boyle CSIRO Livestock Industries (03) 5227 5018

The effect of Newcastle disease vaccination with strain V4 on the course of infections with the Peats Ridge strain of Newcastle disease virus

CSA-18J Dr Peter Daniels CSIRO Livestock Industries (03) 5227 5272

Infectious proventriculitis and stunting syndrome of broiler chickens DAN-171A Dr Rod Reece NSW Dept of Agriculture (02) 4640 6309 Attenuation and characterisation of chicken Eimeria for live vaccines DAQ-259J Dr Wayne Jorgensen Dept of Primary Industries (Qld) (07) 3362 9455 Investigations into the development of a sustainable management strategy for the darkling beetle, Alphitobius diaperinus (Panzer) in broilers

DAQ-273A Dr Trevor Lambkin Dept of Primary Industries (Qld) (07) 3896 9434

Masters in avian health - Dr Rubite MS001-57 Dr Trevor Bagust University of Melbourne (03) 9344 9676 The development of vaccination strategies to control necrotic enteritis in poultry RMI-11A Prof Peter Coloe Royal Melbourne Institute of

Technology (03) 9925 2481

Molecular evaluation of responses to vaccination and challenge by Marek's disease virus RMI-12J Professor Greg Tannock Royal Melbourne Institute of Technology

(03) 9925 3088

Determination of the genomic sequence of Mycoplasma gallisepticum UM-45J Dr Glenn Browning University of Melbourne (03) 8344 7342 Avian Leukosis-J (ALV-J) in Australia: laboratory technologies and research needs UM-49A Dr Trevor Bagust University of Melbourne (03) 9344 9676 Control of intestinal spirochaete infections in chickens UMU-23J A/Prof David Hampson Murdoch University (08) 9360 2287 Control of intestinal spirochaete infections in chickens UMU-29J Professor David Hampson Murdoch University (08) 9360 2287 Effects of organic acids, prebiotics, and enzymes on control of necrotic enteritis and performance of broiler chickens

UNE-75A Dr Mingan Choct University of New England (02) 6773 5121

Typing of Pasteurella multocida UQ-100J A/Professor Linda Blackall University of Queensland (07) 3365 4645 Enhancing mucosal immunity in chickens by novel in-ovo and postnatal vaccination techniques

US-72J Prof Alan Husband University of Sydney (02) 9351 3127


159

Bird Nutrition and Feed Supply Project Title Project No Principal Investigator Organisation Phone No Characterisation of canola meal and cottonseed meal at practical inclusion levels for use in broiler and layer diets

DAQ-264J Dr Rider Perez-Maldonado Dept of Primary Industries (Qld) (07) 3824 3081

Estimating lysine availability by slope-ratio chick assay DAQ-277A Dr Rider Perez-Maldonado Dept of Primary Industries (Qld) (07) 3824 3081 Premium grains for livestock program (stage 2) GRD-3J Dr John Black Grains Research & Development

Corporation (02) 4753 6231

Physiological limitations in energy metabolism reduce production efficiency of broilers SAR-13A Mr Bob Hughes South Australian Research and Development Institute

(08) 8303 7788

Improving the utilisation of dietary amino acids in meat chickens US-80A A/Prof Wayne Bryden University of Sydney (02) 4655 0658 Use of dietary fatty acids to increase protein accretion in broilers US-104A A/Prof Wayne Bryden University of Sydney (02) 4655 0658

Food Safety Project Title Project No Principal Investigator Organisation Phone No Salmonella typing and colonisation of chickens by characterised S. Sofia IMV-3A Dr Michael Heuzenroeder Institute of Medical & Veterinary

Science (08) 8222 3275

Development of campylobacter bio-replacement program and establishment of campylobacter reference centre

UG-3A Dr Victoria Korolik Griffith University (07) 5552 8321

Isolation of genes responsible for Campylobacter jejuni colonisation of the chicken intestinal tract

UG-4A Dr Victoria Korolik Griffith University (07) 5552 8321

Consumer and Community Perceptions Project Title Project No Principal Investigator Organisation Phone No On-farm reduction strategies for Campylobacter spp. DAQ-282A Ms Jeanette Miflin Dept of Primary Industries (Qld) (07) 3362 9520 Chicken meat usage study NMR-1A Ms Carole Yeomans Newspoll Market Research (02) 9281 8100

Animal Welfare Project Title Project No Principal Investigator Organisation Phone No Implementation of the RIRDC broiler welfare audit to industry DAV-185A Prof John Barnett Dept of Natural Resources &

Environment (Vic) (03) 9742 0433

Environmental Management Project Title Project No Principal Investigator Organisation Phone No National environmental management system for the meat chicken industry FSE-1A Mr Eugene McGahan FSA Environmental (07) 4632 8230 Sustainability improvements in the Victorian chicken meat industry (phase 1) JSC-1A Mr Jim Smith James Smith Consulting (03) 9598 8717 Preparation of a database of envrionmental data relevant to chicken meat farms and assessment of odour control strategies

PAE-1A Mr Robin Omerod Pacific Air & Environment Pty Ltd (07) 3844 6400

Reduction of dust emissions from broiler and caged layer sheds SAR-33J Mr Themes Banhazi South Australian Research and Development Institute

(08) 8303 7781


160

RESEARCH TO BE SUPPORTED IN 2001/2002 EGG PROGRAM

Implications of the Changing Economic Environment for the Australian Egg Industry Project Title Project No Principal Investigator Organisation Phone No Options for enhancing industry competitiveness and R&D and marketing efficiency AEI-11A Mr Alan Newton Australian Egg Industry Association (02) 6295 7251

New and Existing Markets Project Title Project No Principal Investigator Organisation Phone No An evaluation of the higher value-added opportunities from the chicken egg DAQ-275A Dr Craig Davis Dept of Primary Industries (Qld) (07) 3406 8611

Public Health Project Title Project No Principal Investigator Organisation Phone No To raise awareness of eggs, cholesterol and health issues AEI-12A Ms Nola Komis Australian Egg Industry Association (02) 9570 9222 Rapid detection of virulent Salmonella in egg and poultry products CIF-1A Dr Jason Wan CRC for International Food

Manufacture and Packaging Science (03) 9742 0320

Eggs with increased arachidonic acid for infant formulas CNR-1A Dr Robert Gibson Child Health Research Institute (08) 8204 5469 Is total egg avoidance really necessary for egg allergy treatment? CNR-2A Dr Maria Makrides Child Health Research Institute (08) 8204 6067 Enriching the iron content of eggs to fulfil niche markets UA-56A Dr Dean Revell University of Adelaide (08) 8303 7911 Egg and egg shell quality control in the Australian egg industry UNE-71A A/Prof Juliet Roberts University of New England (02) 6773 2506

Flock Health and Disease Management Project Title Project No Principal Investigator Organisation Phone No Trialing emergency animal disease arrangements in the Australian egg industry AEI-10A Mr Malcolm Peacock Australian Egg Industry Association (02) 9570 9222 Postgraduate scholarship - Ms Louise Hilton: Therapeutic applications of cytokines in poultry CSA-10J Dr John Lowenthal CSIRO Livestock Industries (03) 5227 5759 Molecular epidemiology of Newcastle disease virus in Australia CSA-11J Dr Allan Gould CSIRO Livestock Industries (03) 5227 5119 Postgradute Scholarship - Jacqueline Kattenbelt: Analysis of virulence determinants of Newcastle disease virus

CSA-13J Dr Allan Gould CSIRO Livestock Industries (03) 5227 5119

Diagnostic tools for differentiation of vvIBDV and characterisation of Australian strains CSA-15J Dr Jagoda Ignjatovic CSIRO Livestock Industries (03) 5227 5769 The effect of Newcastle disease vaccination with strain V4 on the course of infections with the Peats Ridge strain of Newcastle disease virus

CSA-18J Dr Peter Daniels CSIRO Livestock Industries (03) 5227 5272

Efficacy trials of a maternally-delivered recombinant vaccine against coccidiosis CME01-05J Dr Nicholas Smith University of Technology, Sydney (02) 9514 4013 Attenuation and characterisation of chicken Eimeria for live vaccines DAQ-259J Dr Wayne Jorgensen Dept of Primary Industries (Qld) (07) 3362 9455 Studies of cloacal haemorrhage, egg peritonitis, vent trauma and beak trimming in the laying hen

DAV-170A Dr Greg Parkinson Dept of Natural Resources & Environment (Vic)

(03) 9217 4200

Studies of cloacal haemorrhage and beak trimming in the laying hen (II) DAV-188A Dr Greg Parkinson Dept of Natural Resources & Environment (Vic)

(03) 9217 4200


161

Masters in avian health - Dr Rubite MS001-57 Dr Trevor Bagust University of Melbourne (03) 9344 9676 Molecular evaluation of responses to vaccination and challenge by Marek's disease virus RMI-12J Professor Greg Tannock Royal Melbourne Institute of

Technology (03) 9925 3088

Molecular diagnostic tools for wild type and vaccine strains of Marek's disease virus UJC-7A Dr Graham Burgess James Cook University (07) 4781 5472 Determination of the genomic sequence of Mycoplasma gallisepticum UM-45J Dr Glenn Browning University of Melbourne (03) 8344 7342 Postgraduate scholarship - Ms Michelle Peters - Molecular biology of chicken anaemia virus UM-46A Dr Glenn Browning University of Melbourne (03) 8344 7342 Investigating sanitation of surface water for poultry using chlorine - IBDV models UM-51A Dr Trevor Bagust University of Melbourne (03) 9344 9676 Improving mycoplasma vaccines - targets for defined attenuation UM-54A Dr Phillip Markham University of Melbourne (03) 8344 7363 Further development of a live attenuated vaccine for chicken anaemia virus UM-55A Dr Glenn Browning University of Melbourne (03) 8344 7342 Control of intestinal spirochaete infections in chickens UMU-23J A/Prof David Hampson Murdoch University (08) 9360 2287 Control of intestinal spirochaete infections in chickens UMU-29J Prof David Hampson Murdoch University (08) 9360 2287 Effects of diet composition, gut microbial status and feed forms on cannibalism in layers UNE-72A Dr Mingan Choct University of New England (02) 6773 5121 Optimising infectious bronchitis vaccination of laying hens for maximum egg shell quality UNE-76A A/Prof Juliet Roberts University of New England (02) 6773 2506 Typing of Pasteurella multocida UQ-100J A/Prof Linda Blackall University of Queensland (07) 3365 4645 Enhancing mucosal immunity in chickens by novel in-ovo and postnatal vaccination techniques

US-72J Prof Alan Husband University of Sydney (02) 9351 3127

Feed Availability and Nutrition Project Title Project No Principal Investigator Organisation Phone No Characterisation of canola meal and cottonseed meal at practical inclusion levels for use in broiler and layer diets

DAQ-264J Dr Rider Perez-Maldonado Dept of Primary Industries (Qld) (07) 3824 3081

Energy requirements of imported layer strains DAQ-280A Mr David Robinson Dept of Primary Industries (Qld) (07) 3824 3081 Effect of sorghum ergot on the egg chicken industry EGG01-32J Dr John Dingle University of Queensland (07) 5460 1250 Premium grains for livestock program (stage 2) GRD-3J Dr John Black Grains Research & Development

Corporation (02) 4753 6231

Hind gut function in laying hens UNC-12A Dr Robert Taylor University of Newcastle (02) 9872 7203 Inflammatory response to diet in the hindgut of layers UNC-14A Dr Robert Taylor University of Newcastle (02) 9872 7203 Effects of commercial feed enzymes in wheat-based diets on egg and egg shell quality in imported strains of laying hen

UNE-77A A/Prof Juliet Roberts University of New England (02) 6773 2506

Evaluation of Lathyrus cicera as a feed ingredient for layers UWA-61A Dr Colin Hanbury University of Western Australia (08) 9638 3744

Husbandry and Welfare Project Title Project No Principal Investigator Organisation Phone No Modifying egg production systems to meet changing consumer needs DAQ-279A Mr Geofrey Runge Dept of Primary Industries (Qld) (07) 5495 1511 Layer strains for alternative systems DAQ-283A Mr David Robinson Dept of Primary Industries (Qld) (07) 3824 3081 Welfare of laying hens in furnished cages EGG01-21 Dr John Barnett Dept of Natural Resources &

Environment (Vic) (03) 9742 0433

Should claw abrasives be used in cages in Australia? SAR-34A Dr Phil Glatz South Australian Research and Development Institute

(08) 8303 7786

Beak trimming accreditation SAR-35A Dr Phil Glatz South Australian Research and Development Institute

(08) 8303 7786


162

The assessment and development of best management practice techniques for Australian laying hens housed in conventional and alternative laying systems

UQ-93A Prof J Ternouth University of Queensland (07) 5460 1267

Pilot study on the use of time lapse video to study the behaviour of laying hens housed in conventional and modified cages

UQ-97A Mr Geoff Stewart University of Queensland (07) 5460 1417

Non-invasive stress assessment of commercial egg industry practices US-107A Dr Jeff Downing University of Sydney (02) 9351 1600

Husbandry and Welfare Project Title Project No Principal Investigator Organisation Phone No Reduction of dust emissions from broiler and caged layer sheds SAR-33J Mr Themes Banhazi South Australian Research and

Development Institute (08) 8303 7781

Training, Information and Technology Transfer Project Title Project No Principal Investigator Organisation Phone No Identifying communication mediums and issues for the egg industry ALL-1A Ms Vicki Noy Alliance Consulting & Management (07) 3367 1113 National egg industry newsletter DAN-138A Mr Gerry Bolla NSW Dept of Agriculture (02) 4348 1917 Video series - Workplace training for layer farm staff DAN-189A Mr Michael Bourke NSW Dept of Agriculture (02) 6391 3209 National egg industry newsletter DAN-194A Mr Gerry Bolla NSW Dept of Agriculture (02) 4348 1900 Vaccination training manual SAR-36A Dr Phil Glatz South Australian Research and

Development Institute (08) 8303 7786

Chicken Meat and Egg Programs - Agrifutures Australia · Sure-feed Pty Ltd PO Box 1208 WAGGA WAGGA...

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